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• 2015: 9萬元試管嬰兒： 時間：即曰起到10月31日止 報名從速！ https://www.facebook.com/ok7260678/videos/vb.504323362/10153592235483363/?type=2&theater&notif_t=video_processed 看圖秒懂！ 這學術減免計劃是博元婦產科和彰基基因醫學部的共同研究： 1,本研究目的在研究qPCR based PGS 試管嬰兒成功率能提高！ 研究病人的要求： 取卵子數目超過十七個 植入PGS 正常二個胚胎（DET） 計算統計是否如RMANJ一様可達到 懷孕率94.7% ? 活産率84.7% 嗎？ 2. 低劑量陰道黃體素的時間之有效性統計 故有此計劃。 qPCR based PGS 的特殊性：新鮮植入！ 收案數:70位試管嬰兒病人 日期：暫定10月31日 如果未滿七十人則延後收案日期到今年12月31日 研究背景：美國紐澤西的RMANJ團隊 七十三個病人 平均取卵子數目超過十七個 植入qPCR的PGS植入正常二個胚胎DET 計算統計 懷孕率84.7%？ 懷孕率94.7% ? 活産率84.7% ? 　from： F&S 2013。9月 題目：Blastocyst biopsy with comprehensive chromosome screening and fresh embryo transfer significantly increases in vitro fertilization implantation and delivery rates: a randomized controlled trial. 9萬元試管嬰兒 時間：即曰起到10月31日止 報名從速！ 電話:04-7260678 e-mail: ok7260678@gmail.com 博元婦產科9萬2千元"左右"的試管嬰兒明細說明: 1. 為什麼說左右：因為打針過程會有少量劑量上的增減，比如打針不到1950單位，我們會退費；如果是第三天胚胎植入，我們會退費囊胚培養費8000元 2. 9萬2千元左右試管嬰兒服務細節9項如下： 　　整個療程! 　（1）排卵針1950單位 　（2）欣得泰6支 　（3）克得諾1支 　（4）回診抽血6次+超音波4次 　（5）取卵手術 　（6）胚胎培養 　（7）植入胚胎手術 　（8）2週黃體素支持療法 以上為9萬2千元左右試管嬰兒的服務實質內容。 以下為除外條款，若是要做： 1. 胚胎著床前遺傳檢查PGS，另計 2. 胚胎著床前基因檢查PGD費用，另計 3. 冷凍胚胎費用，另計 4. 若要累積胚胎以增加懷孕率，一次試管嬰兒療程算一次的費用，另計 博元婦產科處處關心您 電話：04-7260678 時間：即日起至10月31日止 學術研究背景： 為研究 1. 胚胎快篩一條龍試管嬰兒在增加試管嬰兒成功的效率（若要做PGS，費用另計） 2. 陰道黃體素的LOW DOSE劑量時間之有效性統計 故有此計劃。 電話:04-7260678 e-mail: ok7260678@gmail.com 9萬2試管嬰兒 時間：即曰起到10月31日止 報名從速！ image1.JPG 從我的 iPhone 傳送
• 2015: 《一條龍試管嬰兒qPCR》論文 被Molecular cytogenetics 是一個國際期刊拿來廣告對期刊有好的影響 在這封信裡 發表的論文高居第一名被下載的第一名 發表的qPCR論文 被Molecular cytogenetics 是一個國際 期刊拿來廣告對期刊有好的影響 在這封信裡 發表的論文高居第一名被下載的第一名 七月 發表的論文 被期刊拿來廣告 是最常被下載的論文之一 對期刊有好的影響 在這封信裡 發表的論文高居第一名 http://www.molecular cytogenetics.org/content/8/1/49 http://www.molecularcytogenetics.org/content/8/1/49 內置圖片 1內置圖片 2 內置圖片 4 內置圖片 1內置圖片 3內置圖片 2 ​ BioMed Central news@info.biomedcentral.com 透過 bounce.news.springer.com 21:37 (9 小時前) 寄給 我 Dear Colleague, Below is a selection of the latest, highly accessed articles from the open access journal, Molecular Cytogenetics: - Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification - Partial tetrasomy of the proximal long arm of chromosome 15 in two patients: the significance of the gene dosage in terms of phenotype - Genes on B chromosomes of vertebrates Interested in publishing with BioMed Central? 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Research Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification Yu-Shih Yang1†, Shun-Ping Chang2†, Hsin-Fu Chen13†, Gwo-Chin Ma24†, Wen-Hsiang Lin2, Chi-Fang Lin1, Feng-Po Tsai5, Cheng-Hsuan Wu6, Horng-Der Tsai6, Tsung-Hsien Lee17 and Ming Chen1268* *Corresponding author: Ming Chenmchen_cch@yahoo.com; mingchenmd@gmail.com † Equal contributors Author Affiliations 1 Department of Obstetrics and Gynecology, College of Medicine, National Taiwan University, Taipei, Taiwan 2 Department of Genomic Medicine, and Center for Medical Genetics, Changhua Christian Hospital, Changhua, Taiwan 3 Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei, Taiwan 4 Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, Taiwan 5 Poyuan Women Clinic, Changhua, Taiwan 6 Department of Obstetrics and Gynecology, Changhua Christian Hospital, Changhua, Taiwan 7 Department of Obstetrics and Gynecology, Chung-Shan Medical University, Taichung, Taiwan 8 Department of Life Sciences, Tunghai University, Taichung, Taiwan For all author emails, please log on. Molecular Cytogenetics 2015, 8:49 doi:10.1186/s13039-015-0140-9 Yu-Shih Yang, Shun-Ping Chang, Hsin-Fu Chen and Gwo-Chin Ma contributed equally to this work. The electronic version of this article is the complete one and can be found online at: http://www.molecularcytogenetics.org/content/8/1/49 Received: 11 February 2015 Accepted: 7 May 2015 Published: 8 July 2015 © 2015 Yang et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Abstract Background Aneuploidy is an important etiology of implantation failure and quantitative real-time polymerase chain reaction (qPCR) seems a promising preimplantation genetic screening (PGS) technology to detect aneuploidies. This verification study aimed at verifying the impact on reproductive outcomes in in vitrofertilization (IVF) cycles using fresh embryo transfer (FET) in which the embryos were selected by blastocyst biopsy with qPCR-based PGS in our settings. Results A total of 13 infertile couples with more than once failed in vitro fertilization were enrolled during July to October of 2014. PGS was conducted by qPCR with selectively amplified markers to detect common aneuploidies (chromosomes 13, 18, 21, X, and Y). The design of the qPCR molecular markers adopted the locked nucleic acid (LNA) strategy. The blastocyst biopsy was performed on Day 5/6 and the PGS was done on the same day, which enabled FET. A total of 72 blastocysts were biopsied. Successful diagnoses were established in all embryos and the rate of successful diagnosis was 100 %. The aneuploidy rate was 38.9 % (28/72). 28 embryos were transferred. The clinical pregnancy rate was 61.5 % (8/13) per cycle. Early first trimester abortion was encountered in 1 and the ongoing pregnancy rate was 53.8 % (7/13) per cycle. Conclusion This study verified the favorable outcome of adopting PGS with qPCR + FET in our own setting. Expanding the repertoire of aneuploidies being investigated (from a limited set to all 24 chromosomes) is underway and a randomized study by comparing qPCR and other PGS technologies is warranted. Keywords: Aneuploidy; Blastocyst; Fresh embryo transfer; PGS; qPCR http://www.molecularcytogenetics.org/content/8/1/49 Research Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification Yu-Shih Yang1†, Shun-Ping Chang2†, Hsin-Fu Chen13†, Gwo-Chin Ma24†, Wen-Hsiang Lin2, Chi-Fang Lin1, Feng-Po Tsai5, Cheng-Hsuan Wu6, Horng-Der Tsai6, Tsung-Hsien Lee17 and Ming Chen1268* *Corresponding author: Ming Chenmchen_cch@yahoo.com; mingchenmd@gmail.com † Equal contributors Author Affiliations 1 Department of Obstetrics and Gynecology, College of Medicine, National Taiwan University, Taipei, Taiwan 2 Department of Genomic Medicine, and Center for Medical Genetics, Changhua Christian Hospital, Changhua, Taiwan 3 Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei, Taiwan 4 Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, Taiwan 5 Poyuan Women Clinic, Changhua, Taiwan 6 Department of Obstetrics and Gynecology, Changhua Christian Hospital, Changhua, Taiwan 7 Department of Obstetrics and Gynecology, Chung-Shan Medical University, Taichung, Taiwan 8 Department of Life Sciences, Tunghai University, Taichung, Taiwan For all author emails, please log on. Molecular Cytogenetics 2015, 8:49 doi:10.1186/s13039-015-0140-9 Yu-Shih Yang, Shun-Ping Chang, Hsin-Fu Chen and Gwo-Chin Ma contributed equally to this work. The electronic version of this article is the complete one and can be found online at: http://www.molecularcytogenetics.org/content/8/1/49 Received: 11 February 2015 Accepted: 7 May 2015 Published: 8 July 2015 © 2015 Yang et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Abstract Background Aneuploidy is an important etiology of implantation failure and quantitative real-time polymerase chain reaction (qPCR) seems a promising preimplantation genetic screening (PGS) technology to detect aneuploidies. This verification study aimed at verifying the impact on reproductive outcomes in in vitrofertilization (IVF) cycles using fresh embryo transfer (FET) in which the embryos were selected by blastocyst biopsy with qPCR-based PGS in our settings. Results A total of 13 infertile couples with more than once failed in vitro fertilization were enrolled during July to October of 2014. PGS was conducted by qPCR with selectively amplified markers to detect common aneuploidies (chromosomes 13, 18, 21, X, and Y). The design of the qPCR molecular markers adopted the locked nucleic acid (LNA) strategy. The blastocyst biopsy was performed on Day 5/6 and the PGS was done on the same day, which enabled FET. A total of 72 blastocysts were biopsied. Successful diagnoses were established in all embryos and the rate of successful diagnosis was 100 %. The aneuploidy rate was 38.9 % (28/72). 28 embryos were transferred. The clinical pregnancy rate was 61.5 % (8/13) per cycle. Early first trimester abortion was encountered in 1 and the ongoing pregnancy rate was 53.8 % (7/13) per cycle. Conclusion This study verified the favorable outcome of adopting PGS with qPCR + FET in our own setting. Expanding the repertoire of aneuploidies being investigated (from a limited set to all 24 chromosomes) is underway and a randomized study by comparing qPCR and other PGS technologies is warranted. Keywords: Aneuploidy; Blastocyst; Fresh embryo transfer; PGS; qPCR Background Preimplantation genetic screening (PGS) by Day 3 cleavage stage embryo biopsy followed by examination with fluorescence in situ hybridization (FISH) was once popular strategy for women with advanced maternal age (AMA) [1]. This strategy was based upon an assumption that aneuploidy, which is associate with AMA, is an important factor for implantation failure and therefore such strategy can improve the reproductive outcome [2]. However, a famous randomized study published by Mastenbroek and colleagues reported that such FISH-based PGS with Day 3 biopsy did not confer advantages in women with AMA regarding the live-birth rates [3], and it is then called the “Mastenbroek controversy”. This unexpected finding was subsequently proved by many randomized controlled trials [4] except in one series [5]. Some other researchers thus proposed different strategies including adopting different timing of biopsy (trophectoderm biopsy versus cleavage-stage blastomere biopsy), different molecular technologies which can detect all 24 chromosomes instead of only a few selected chromosomes (which is a major limitation of FISH), and different timing of embryo transfer (fresh or frozen) [1], [6], [7]. Mounting evidences had suggested blastomere biopsy at Day 3 cleavage stage embryos does impair the implantation potential whereas trophectoderm biopsy at Day 5/6 blastocyst does not [8]–[10]. Meanwhile, it is straightforward that using more sophisticated molecular tools may select a better or “more normal” embryo. Competing technologies include array comparative genomic hybridization (CGH), single nucleotide polymorphism (SNP) array, quantitative real-time polymerase chain reaction (qPCR), and next generation sequencing (NGS) [11]. Amongst them, NGS remains to be mostly investigational because of the resources and cost of bioinformatics analyses [12], [13], and therefore the most feasible tools at the present seem to be qPCR and array-based technologies, including SNP array and array CGH [8], [14]–[18]. It is now known that mosaicism is a common phenomenon in early human embryo development, and therefore how to select the embryos with best implantation potential becomes a vital question [19]. It is shown by the research groups who proposed for qPCR that qPCR is superior to array CGH since it enjoys a lower false-positive rate for aneuploidy detection (0 % for qPCR and 5.4 % for array CGH in 122 embryos analyzed), and thereby preserving more embryos available for transfer and ensues a better reproductive outcome [14]. There are now commercially available PGS platforms using array CGH in the market (such as those produced by BlueGnome, UK) and has become very popular because of the feasibility of an easy-to-use system, especially for in vitro fertilization (IVF) centers without a genetic lab as a supporting resort [11]. However, since the outcomes achieved by the team proposing qPCR are compellingly excellent. It thus poses stress upon researchers of the similar field to verify qPCR as a replacement or an alternative to the current popular PGS by array CGH platforms [14]. In this small verification study, we aimed to assess the PGS by qPCR strategy coupled with trophectoderm biopsy at Day 5/6 blastocyst stage. In the initial stage we first verified qPCR only at selected common aneuploidies including chromosome 13, 18, 21, X, and Y (these aneuploidies can be confirmed by FISH). The reasons we assessed only selected chromosomes are three: First, the detail design and experimental conditions regarding qPCR on all 24 chromosomes are patent protected and we need staged efforts to devise and validate our own. Therefore, in this study we adopted a method called locked nucleic acid (LNA) technology to design our qPCR platform [20]; Second, for a rapid overnight diagnosis which is warranted for fresh embryo transfer (FET), it takes a qPCR machine with a 384-well for only one embryo [18], which may not be feasible in real settings in most genetic labs; Third, we believe the “Mastenbroek controversy” may result not only from lacking an effective genotyping system to screen a “better” embryo, but also from the proven impairment of the implantation potential when biopsy at Day 3 cleavage stage embryos [3], [4], [6]. The team proposing qPCR in one hand arguing qPCR is better than FISH because it can examine all 24 chromosomes but in the other hand claiming despite array CGH should have a better resolution than qPCR whereas implying array CGH is “too sensitive” since it has a more false-positive rate when SNP array is used as a gold standard [14], [16], [18]. We thus are interested to know if it is feasible by using a limited qPCR strategy with examination of only selected common aneuploidies (similar to FISH) coupled with trophectoderm biopsy of Day 5/6 blastocyst stage embryos can still achieve a favorable reproductive outcome. It is ethical since it is by theory better than the trial protocol reported in the “Mastenbroek controversy” trial [3], in which a similar number of aneuploidies was assessed by FISH but the biopsy timing was set at Day 3 (when cleavage-stage embryos), a timing now being recognized to be inferior to Day 5/6 blastocyst stage regarding the implantation potential [9]. After the feasibility is confirmed in our setting, it is then justified to expand the repertoire of aneuploidies to 9 chromosomes (13, 15, 16, 17, 18, 21, 22, X and Y. See Rubio et al., 2013 a[5]), or 11 chromosomes (7, 9, 11, 13, 14, 15, 18, 21, 22, X and Y. These are the most commonly found trisomies at abortus. See Wang et al., 2014 [21]) and eventually to 24 chromosomes (1–22, X and Y. See Treff et al., 2012 [1]) in the future. Results Pre-clinical validation Pre-clinical validation for the PGS by qPCR was performed on 54 surplus frozen embryos (Additional file 1: Table S1). Fifty-three of these embryos were successfully diagnosed and the successful diagnostic rate was 98.1 % (53/54). Twenty-four embryos were found to carry aneuploidy on tested chromosomes and the rate of aneuploidy was thus 44.4 % (24/53). The only embryo failed to provide a confirmed diagnosis showed poor qPCR signal, possibly due to a low amount of or degraded DNA in the biopsied sample. FISH diagnosis for that embryo was negative for numerical disorders involving chromosome 13, 18, 21, X, and Y. The remaining cells of this biopsied embryo were sent for array CGH and the diagnosis was trisomy 22. Clinical verification A total of 72 embryos at Day 5/6 blastocyst stage from 13 patients were biopsied and assessed, and all (100 %) of them were successfully diagnosed (Table 1 and Additional file 1: Table S1). Twenty-eight (38.9 %) of these embryos were aneuploid, which included 4 monosomy 13, 5 monosomy 21, 7 trisomy 13, 1 trisomy 18, 9 trisomy 21, 1 monosomy 13 + trisomy 18, and 1 monosomy 18 + monosomy 21. The remaining 44 (61.1 %) embryos were euploidy, of which 28 were transferred into the 13 patients in 13 cycles. The average number of the embryos being transferred is 2.15, actually only 1–2 embryos were transferred in all women of the Group B (non-AMA group, n = 10). The only 2 women who were transferred for 4 embryos are both in the Group A (AMA group, n = 3). The transfer procedure follows the medical rule of Taiwan that the upper limit of the number of the embryos being transferred is 4. These transfers achieved 8 pregnancies, and therefore the clinical pregnancy rate was 61.5 % (8/13) per cycle. However, one early abortion was noted and the ongoing pregnancy rate up to the second trimester was 53.8 % (7/13) per transfer cycle. There were no monozygotic twinning and a total of 11 sacs were noted. Three women who were transferred for 2 embryos in the Group B had singleton pregnancies, and two women who were transferred for 2 embryos in the Group B and 2 women who were transferred for 4 embryos in the Group A had dizygotic twin pregnancies (Additional file 1: Table S1). The sustained implantation rate was thus 39.3 % (11/28) per transferred fresh embryo (Table 1). Totally, there are 2 of 3 women in Group A had ongoing pregnancies. Six women in Group B had clinical pregnancies, and 5 of them had sustained ongoing pregnancies. There are no statistical significant differences noted between these two groups regarding aneuploidy (Chi square test, p = 0.094), clinical pregnancy (Fisher's exact test, p = 1), and ongoing pregnancy (Fisher's exact test, p = 1). Array CGH for the only abortion case revealed the karyotype was 46,XY. Table 1. Clinical outcomes of PGS by qPCR with fresh embryo transfer (FET) verification series Discussion The major difficulty of comprehensive chromosome screening (CCS) by qPCR for PGS is to devise a primer set to successfully amplify the molecular markers being selected in a single experiment by optimizing the conditions for melting temperatures and experimental time intervals. In this study we successfully adopted a smart design called LNA methodology [20], [22]–[24] for qPCR PGS and verified the strategy by achieving a favorable ongoing pregnancy rate of 53.8 %. We first validated the genotyping platform we devised by achieving an almost 100 % successful diagnosis rate (98.1 %), and then verified the whole protocol of qPCR PGS followed by FET. Since the sample size is small (n = 13 cycles), it is not surprising that we failed to observe significant difference regarding the aneuploidy rates between the AMA (n = 3) and the non-AMA group (n = 10) in our patients. Meanwhile, the aneuploidy rate in our series (38.9 %) is apparently much lower than the rates observed by 24-chromosome qPCR or array CGH (more than 60 %), which is obviously due to the fact that we only selected a limited set of common aneuploidies that can ensue live births in humans [14], [15]. It has been a routine practice in our settings that PGS by FISH at Day 3 cleavage-stage embryos since 2005 and it is ethical for us to offer our patients who opted for this alternative protocol a strategy which has an advantage of an established less detrimental effect upon implantation potential by changing the timing of biopsy from Day 3 to Day 5/6 and the same repertoire of common aneuploidies being tested (but simply using qPCR instead of FISH) to enable FET[9]. The reason only trisomy 13, 18, 21, and numerical disorders involving X and Y were tested in our settings is because these are the only aneuploidies that may result in live births in humans. For people who opted for this strategy and came to our clinics, their major concern is more focused at aneuploidies instead of live birth rates. In addition, it is well recognized that despite fewer embryos would be available for transfer if being cultured in vitro onto the blastocyst stage when compared with the cleavage-stage embryos, the disadvantage may be offset by the fact that the aneuploidy rate of the blastocysts is much lower than of the cleavage-stage embryos. An observational study had reported that karyotypic evolution did exist in early human embryogenesis, in which some cleavage stage embryos classified as aneuploidy by FISH eventually developed into euploid blastocysts if by SNP array[25]. A recent randomized study even pointed out that FISH-based PGS conferred advantages at women who are in the AMA group, but not in those with repeated implantation failures group, as long as the biopsy was performed at the blastocyst stage [5]. A recent randomized trial (the BEST trial) had demonstrated that qPCR PGS CCS followed by single embryo transfer (SET) can enhance the feasibility of SET by selecting a single euploid embryo with high reproductive potential without compromising the delivery rates when compared with transfer of two unselected embryos, which instead resulted in more multiple pregnancies and the associated complications such as preterm delivery, low birth weight, and NICU (Neonatal Intensive Care Unit) admission [26]. We admit our pilot verification study had suffered from the fact that only common aneuploidies of five chromosome pairs (13, 18, 21, X, and Y) were screened and apparently it is the reason that the implantation rate per embryo in our study (39.3 %) is much lower than that in the BEST trial (69 %) [26]. However, our patients were all informed consented and knew the disadvantage before joining the study, and the limitations of the local setting were clearly disclosed to the patients. The patients were free to choose another PGS protocol by array CGH (Option 2, please refer to Patients and methods section) instead. Conclusions The merit of our work is that this is the first effort from research groups other than the original group who proposed the qPCR CCS (Reproductive Medicine Associates of New Jersey, Morristown, NJ, USA) trying to validate and verify this novel genotyping platform which was developed in-house by ourselves. Despite our results did not have matched controls such as those cycles receiving IVF but without PGS, the ongoing pregnancy rate is similar to a previous PGD series from our setting (53.8 % versus 50 %, please refer to Chang et al., 2013 [27]), indicating this protocol is clinically feasible. Future efforts will be made upon expanding the repertoire of the PGS by qPCR CCS to all 24 chromosomes, as well as carefully-designed randomized controlled trials to compare this genotyping platform with others (including no-screening, array CGH, and NGS). Patients and methods Design and selection of molecular markers for qPCR PGS An in-house screening system by dual-color qPCR was developed with specific primers for the targeted loci (situated at chromosome 13, 18, 21, X, and Y) and one internal control locus (situated at chromosome 1, the reason we chose chromosome 1 as control is because it is the least found trisomy in humans, see Wang et al., 2014 [21]). A total of 16 targeted loci and one control locus were selected. For the detailed information about the targeted loci, please refer to Table 2. Specific primers for amplification of each locus were designed by using the software Oligo 6.71 (Molecular Biology Insights, Colorado, USA), and all primer sets are flanked on each side of the exon-intron boundary to avoid possible mRNA interference. The applications of LNA-modified probes are well-established for quantifying levels of gene expression with an advantage of reducing complexity [22], [24], but has not been applied to the detection of chromosomal copy number alterations. Two kinds of LNA-modified probes, labeled with FAM™ and HEX™ (Integrated DNA Technologies, Iowa, USA) at the 5’-end, were used for quantifying the genomic copy numbers of targeted loci and control. Table 2. Summary of the targeted regions for screening of common aneuploidies Validation of qPCR in detecting common aneuploidies Each 5 cells separated from the cell lines of known common aneuploidies (including trisomy 21, trisomy 13, trisomy 18, 47,XXY, and 45,X) were processed by cell lysis of proteinase K, and the products were subjected to a 50-μL reaction volume of multiplex nested-PCR amplication for 18 cycles using an Applied Biosystems Veriti thermal cycler (Life Technologies, California, USA), and then the PCR products were purified using Agencourt AMPure XP system (Beckman Coulter, California, USA). Dual color hydrolysis probe assays were performed in triplicate to normalize and simpify calculation and to evaluate chromosomal copies, using Lightcycler 480 probes Master (Roche, Mannheim, Germany), a 20-μL reaction volume, a 96-well plate, and a 7 Light Cycler 480 Real-Time PCR System, as recommended by the supplier (Roche, Mannheim, Germany). Each well contains a particular target, and a common control reaction. A unique method of the standard delta delta threshold cycle (ΔΔCp) method was used for relative quantification. In our experiments, the Cp variation of all HEX™ reactions obtained for each well of the same sample will be controlled and ranged in less than 0.2, indicating the test sample was evenly distributed to each well. Each chromosome-specific ΔCp was calculated from the Cp of the FAM™ reactions targeting a specific chromosome minus the control Cp of the HEX™ reactions targeting the chromosome 1 within the same well. The same process was applied to individually determine the ΔCp for each targeted chromosome of the test sample, including reference set of normal male cell lines [BCRC number: 08C0011, 08C0012, 08C0013, 08C0021 and 08C0025]. Each chromosome- specific ΔCp was then normalized to the average chromosome-specific ΔCp values derived from the same evaluation of the reference set, which had been confirmed by FISH method. The calibrated chromosome-specific ΔCp values were used to calculate fold change by considering the ΔΔCp values as the negative exponent of 2, as previously described [18], [23]. The methodology was designed to specifically identify whole-chromosome but not segmental aneuploidy. The flowchart and diagram of the in-house qPCR PGS system were illustrated in Fig. 1. This qPCR was capable of accurate aneuploidy screening in 4 h, which allowed rapid evaluation of the trophectoderm biopsies and therefore provided a feasible opportunity for subsequent FET. thumbnailFig. 1. The diagram of the in-house qPCR PGS system. The flowchart of (a) detection of common aneuploidies, and (b) signal normalization and data analysis Pre-clinical validation upon surplus frozen embryos Fifty-four thawed frozen embryos at blastocyst stage were biopsied and sent for qPCR and FISH analyses. These embryos were retrieved from the surplus frozen embryos of couples who already conceived and would be discarded if not being investigated for research purpose. In those embryos diagnosed with common chromosome aneuploidies, array CGH was used to confirm the diagnoses. Clinical verification for fresh embryos In our setting we used to offer two PGS protocols. One option (Option 1) is Day 3 biopsy followed by PGS with FISH and FET; the other option (Option 2) is Day 5/6 trophectoderm biopsy followed by PGS with array CGH and frozen embryo transfer (because array CGH takes time and FET was not feasible at the study period). Only patients who had history of failed IVF (without PGS) for at least once and who opted for Option 1 were given the chance of joining this study as an alternative. All patients chose to join this study were informed consented and their autonomy was fully respected. They could choose to withdraw from the study at any time during the study period and were fully aware of the alternatives, including sticking to Option 1 or instead chose Option 2 without joining the study. During July to October of 2014, 13 infertile couples were enrolled. Among these 13 patients, 3 of them had AMA (37, 43, 45 years old respectively). The mean age of the total 13 patients was 34.1 years. No confounding factors that may affect implantation such as immune aberrations (antiphospholipid antibody syndrome in the mother), balanced translocation carriers (in both couples), and thrombophilias (protein C/S/antithrombin III deficiency) existed in these couples when they were enrolled. Clinical pregnancy was defined as positive urine HCG. Ongoing pregnancy rate (per cycle) was defined as those pregnancies proceeding into second trimester (and for each embryo being transferred, sustained implantation rate was used). 13 couples were classified as those whose age is older or equal to 35 years (Group A), and those whose age is less than 35 years (Group B). Chi square test or Fisher’s exact test was used to compare the reproductive outcomes between the groups regarding the rate of aneuploidy, clinical pregnancy, and ongoing pregnancy. Notably according to the regulations of Taiwan government, the patients would not know the results of the fetal sex. Even in aneuploidies involving sex chromosomes (specifically 47,XXY and 45,X), the results would not be disclosed to patients and only “aneuploidy” would be told to the patients and the aneuploid embryos would not be selected for transfer. In those embryos diagnosed to have chromosomal aneuploidies by qPCR, array CGH was used to confirm the diagnoses. Abbreviations AMA: Advanced maternal age CCS: Comprehensive chromosome screening CGH: Comparative genomic hybridization FET: Fresh embryo transfer FISH: Fluorescence in situ hybridization IVF: in vitro fertilization LNA: Locked nucleic acid NGS: Next generation sequencing PGS: Preimplantation genetic screening qPCR: Quantitative real-time polymerase chain reaction SET: Single embryo transfer SNP: Single nucleotide polymorphism Competing interests The authors declare that they have no competing interests. Authors’ contributions YSY, HFC, SPC, FPT, MC designed the study. SPC, GCM, MC developed the novel in-house genotyping platform. SPC, GCM, WHL, CFL, MC did the experiments. YSY, HFC, FPT, CHW, HDT, MC recruited the patients. YSY, HFC, GCM, THL, MC analyzed the results. YSY, SPC, GCM, WHL, MC wrote the paper. All authors read and approved the final manuscript. Additional file Additional file 1: Table S1.. Summary of the pre-clinical validation (for 54 surplus frozen embryos) and clinical verification (for 13 patients with 54 embryos) of PGS by qPCR. Format: DOCX Size: 32KB Download fileOpen Data Acknowledgements This study is partly funded by a grant from Changhua Christian Hospital, Taiwan to Ming Chen (103-CCH-IST-006). The authors are grateful for the genuine support and advice from Dr. Shou-Jen Kuo, the Superintendent of Changhua Christian Hospital, who also participated in the development of the novel in-house qPCR CCS system. References Treff NR, Scott RT. Methods for comprehensive chromosome screening of oocytes and embryos: capabilities, limitations, and evidence of validity. 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Preimplantation genetic screening using fluorescence in situ hybridization in patients with repetitive implantation failure and advanced maternal age: two randomized trials. Fertil Steril. 2013;99(5):1400–7.a Braude P. Selecting the 'best' embryos: prospects for improvement. Reprod Biomed Online. 2013; 27(6):244-53.Publisher Full Text OpenURL Mastenbroek S, Repping S. Preimplantation genetic screening: back to the future. Hum Reprod. 2014; 29(9):1846-50. PubMed Abstract |Publisher Full Text OpenURL Scott RT, Ferry K, Su J, Tao X, Scott K, Treff NR. Comprehensive chromosome screening is highly predictive of the reproductive potential of human embryos: a prospective, blinded, nonselection study. Fertil Steril. 2012; 97(4):870-5. PubMed Abstract |Publisher Full Text OpenURL Scott KL, Hong KH, Scott RT. Selecting the optimal time to perform biopsy for preimplantation genetic testing. Fertil Steril. 2013;100(3):608-14. PubMed Abstract | Publisher Full Text OpenURL Scott RT, Upham KM, Forman EJ, Hong KH, Scott KL, Taylor D et al..Blastocyst biopsy with comprehensive chromosome screening and fresh embryo transfer significantly increases in vitro fertilization implantation and delivery rates: a randomized controlled trial.Fertil Steril. 2013; 100(3):697-703. PubMed Abstract |Publisher Full Text OpenURL Handyside AH. 24-chromosome copy number analysis: a comparison of available technologies. Fertil Steril. 2013;100(3):595-602. PubMed Abstract | Publisher Full Text OpenURL Fiorentino F, Bono S, Biricik A, Nuccitelli A, Cotroneo E, Cottone G et al..Application of next-generation sequencing technology for comprehensive aneuploidy screening of blastocysts in clinical preimplantation genetic screening cycles. Hum Reprod. 2014;29(12):2802-13. PubMed Abstract | Publisher Full Text OpenURL Wells D, Kaur K, Grifo J, Glassner M, Taylor JC, Fragouli E et al.. Clinical utilisation of a rapid low-pass whole genome sequencing technique for the diagnosis of aneuploidy in human embryos prior to implantation. J Med Genet. 2014; 51(8):553-62. PubMed Abstract |Publisher Full Text OpenURL Capalbo A, Treff NR, Cimadomo D, Tao X, Upham K, Ubaldi FM, et al. Comparison of array comparative genomic hybridization and quantitative real-time PCR-based aneuploidy screening of blastocyst biopsies. Eu J Hum Genet. 2014;doi:10.1038/ejhg.2014.222(in press). Rubio C, Rhodrigo L, Mir P, Mateu E, Peinado V, Milan M, et al. Use of array comparative genomic hybridization (array-CGH) for embryo assessment: clinical results. Fertil Steril. 2013;99(4):1044–8.b Schoolcraft WB, Treff NR, Stevens JM, Ferry K, Katz-Jaffe M, Scott RT.Live birth outcome with trophectoderm biopsy, blastocyst vitrification, and single-nucleotide polymorphism microarray-based comprehensive chromosome screening in infertile patients.Fertil Steril. 2011; 96(3):638-40. PubMed Abstract | Publisher Full Text OpenURL Treff NR, Levy B, Su J, Northrop LE, Tao X, Scott RT. SNP microarray-based 24 chromosome aneuploidy screening is significantly more consistent than FISH. Mol Hum Reprod. 2010; 16(8):583-9.PubMed Abstract | Publisher Full Text OpenURL Treff NR, Tao X, Ferry KM, Su J, Taylor D, Scott RT. Development and validation of an accurate quantitative real-time polymerase chain reaction-based assay for human blastocyst comprehensive chromosomal aneuploidy screening. Fertil Steril. 2012; 97(4):819-24. PubMed Abstract | Publisher Full Text OpenURL Vanneste E, Voet T, Le Caignec C, Ampe M, Konings P, Melotte C et al..Chromosome instability is common in human cleavage-stage embryos. Nat Med. 2009; 15(5):577-83. PubMed Abstract |Publisher Full Text OpenURL Brunet E, Corgnali M, Cannata F, Perrouault L, Giovannangeli C.Targeting chromosomal sites with locked nucleic acid-modified triplex-forming oligonucleotides: study of efficiency dependence on DNA nuclear environment. Nucleic Acids Res. 2006; 34(16):4546-53. PubMed Abstract | Publisher Full Text OpenURL Wang BT, Chong TP, Boyar FZ, Kopita KA, Ross LP, El-Naggar MM et al..Abnormalities in spontaneous abortions detected by G-banding and chromosomal microarray analysis (CMA) at a national reference laboratory. Mol Cytogenet. 2014; 22:7-33. OpenURL Mouritzen P, Nielsen PS, Jacobsen N, Noerholmn M, Lomholt C, Pfundheller HM et al.. The ProbeLibrary—expression profiling 99 % of all human genes usingonly 90 dual-labeled real-time PCR probes. Biotechniques. 2004; 37:492-495. OpenURL Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative CT method. 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• 2015: 發表的 《一條龍試管嬰兒qPCR》論文 被Molecular cytogenetics 是一個國際期刊拿來廣告對期刊有好的影響 在這封信裡 發表的論文高居第一名被下載的第一名 發表的qPCR論文 被Molecular cytogenetics 是一個國際 期刊拿來廣告對期刊有好的影響 在這封信裡 發表的論文高居第一名被下載的第一名 七月 發表的論文 被期刊拿來廣告 是最常被下載的論文之一 對期刊有好的影響 在這封信裡 發表的論文高居第一名 http://www.molecular cytogenetics.org/content/8/1/49 http://www.molecularcytogenetics.org/content/8/1/49 內置圖片 1內置圖片 2 內置圖片 4 內置圖片 1內置圖片 3內置圖片 2 ​ BioMed Central news@info.biomedcentral.com 透過 bounce.news.springer.com 21:37 (9 小時前) 寄給 我 Dear Colleague, Below is a selection of the latest, highly accessed articles from the open access journal, Molecular Cytogenetics: - Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification - Partial tetrasomy of the proximal long arm of chromosome 15 in two patients: the significance of the gene dosage in terms of phenotype - Genes on B chromosomes of vertebrates Interested in publishing with BioMed Central? 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Research Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification Yu-Shih Yang1†, Shun-Ping Chang2†, Hsin-Fu Chen13†, Gwo-Chin Ma24†, Wen-Hsiang Lin2, Chi-Fang Lin1, Feng-Po Tsai5, Cheng-Hsuan Wu6, Horng-Der Tsai6, Tsung-Hsien Lee17 and Ming Chen1268* *Corresponding author: Ming Chenmchen_cch@yahoo.com; mingchenmd@gmail.com † Equal contributors Author Affiliations 1 Department of Obstetrics and Gynecology, College of Medicine, National Taiwan University, Taipei, Taiwan 2 Department of Genomic Medicine, and Center for Medical Genetics, Changhua Christian Hospital, Changhua, Taiwan 3 Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei, Taiwan 4 Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, Taiwan 5 Poyuan Women Clinic, Changhua, Taiwan 6 Department of Obstetrics and Gynecology, Changhua Christian Hospital, Changhua, Taiwan 7 Department of Obstetrics and Gynecology, Chung-Shan Medical University, Taichung, Taiwan 8 Department of Life Sciences, Tunghai University, Taichung, Taiwan For all author emails, please log on. Molecular Cytogenetics 2015, 8:49 doi:10.1186/s13039-015-0140-9 Yu-Shih Yang, Shun-Ping Chang, Hsin-Fu Chen and Gwo-Chin Ma contributed equally to this work. The electronic version of this article is the complete one and can be found online at: http://www.molecularcytogenetics.org/content/8/1/49 Received: 11 February 2015 Accepted: 7 May 2015 Published: 8 July 2015 © 2015 Yang et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Abstract Background Aneuploidy is an important etiology of implantation failure and quantitative real-time polymerase chain reaction (qPCR) seems a promising preimplantation genetic screening (PGS) technology to detect aneuploidies. This verification study aimed at verifying the impact on reproductive outcomes in in vitrofertilization (IVF) cycles using fresh embryo transfer (FET) in which the embryos were selected by blastocyst biopsy with qPCR-based PGS in our settings. Results A total of 13 infertile couples with more than once failed in vitro fertilization were enrolled during July to October of 2014. PGS was conducted by qPCR with selectively amplified markers to detect common aneuploidies (chromosomes 13, 18, 21, X, and Y). The design of the qPCR molecular markers adopted the locked nucleic acid (LNA) strategy. The blastocyst biopsy was performed on Day 5/6 and the PGS was done on the same day, which enabled FET. A total of 72 blastocysts were biopsied. Successful diagnoses were established in all embryos and the rate of successful diagnosis was 100 %. The aneuploidy rate was 38.9 % (28/72). 28 embryos were transferred. The clinical pregnancy rate was 61.5 % (8/13) per cycle. Early first trimester abortion was encountered in 1 and the ongoing pregnancy rate was 53.8 % (7/13) per cycle. Conclusion This study verified the favorable outcome of adopting PGS with qPCR + FET in our own setting. Expanding the repertoire of aneuploidies being investigated (from a limited set to all 24 chromosomes) is underway and a randomized study by comparing qPCR and other PGS technologies is warranted. Keywords: Aneuploidy; Blastocyst; Fresh embryo transfer; PGS; qPCR http://www.molecularcytogenetics.org/content/8/1/49 Research Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification Yu-Shih Yang1†, Shun-Ping Chang2†, Hsin-Fu Chen13†, Gwo-Chin Ma24†, Wen-Hsiang Lin2, Chi-Fang Lin1, Feng-Po Tsai5, Cheng-Hsuan Wu6, Horng-Der Tsai6, Tsung-Hsien Lee17 and Ming Chen1268* *Corresponding author: Ming Chenmchen_cch@yahoo.com; mingchenmd@gmail.com † Equal contributors Author Affiliations 1 Department of Obstetrics and Gynecology, College of Medicine, National Taiwan University, Taipei, Taiwan 2 Department of Genomic Medicine, and Center for Medical Genetics, Changhua Christian Hospital, Changhua, Taiwan 3 Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei, Taiwan 4 Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, Taiwan 5 Poyuan Women Clinic, Changhua, Taiwan 6 Department of Obstetrics and Gynecology, Changhua Christian Hospital, Changhua, Taiwan 7 Department of Obstetrics and Gynecology, Chung-Shan Medical University, Taichung, Taiwan 8 Department of Life Sciences, Tunghai University, Taichung, Taiwan For all author emails, please log on. Molecular Cytogenetics 2015, 8:49 doi:10.1186/s13039-015-0140-9 Yu-Shih Yang, Shun-Ping Chang, Hsin-Fu Chen and Gwo-Chin Ma contributed equally to this work. The electronic version of this article is the complete one and can be found online at: http://www.molecularcytogenetics.org/content/8/1/49 Received: 11 February 2015 Accepted: 7 May 2015 Published: 8 July 2015 © 2015 Yang et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Abstract Background Aneuploidy is an important etiology of implantation failure and quantitative real-time polymerase chain reaction (qPCR) seems a promising preimplantation genetic screening (PGS) technology to detect aneuploidies. This verification study aimed at verifying the impact on reproductive outcomes in in vitrofertilization (IVF) cycles using fresh embryo transfer (FET) in which the embryos were selected by blastocyst biopsy with qPCR-based PGS in our settings. Results A total of 13 infertile couples with more than once failed in vitro fertilization were enrolled during July to October of 2014. PGS was conducted by qPCR with selectively amplified markers to detect common aneuploidies (chromosomes 13, 18, 21, X, and Y). The design of the qPCR molecular markers adopted the locked nucleic acid (LNA) strategy. The blastocyst biopsy was performed on Day 5/6 and the PGS was done on the same day, which enabled FET. A total of 72 blastocysts were biopsied. Successful diagnoses were established in all embryos and the rate of successful diagnosis was 100 %. The aneuploidy rate was 38.9 % (28/72). 28 embryos were transferred. The clinical pregnancy rate was 61.5 % (8/13) per cycle. Early first trimester abortion was encountered in 1 and the ongoing pregnancy rate was 53.8 % (7/13) per cycle. Conclusion This study verified the favorable outcome of adopting PGS with qPCR + FET in our own setting. Expanding the repertoire of aneuploidies being investigated (from a limited set to all 24 chromosomes) is underway and a randomized study by comparing qPCR and other PGS technologies is warranted. Keywords: Aneuploidy; Blastocyst; Fresh embryo transfer; PGS; qPCR Background Preimplantation genetic screening (PGS) by Day 3 cleavage stage embryo biopsy followed by examination with fluorescence in situ hybridization (FISH) was once popular strategy for women with advanced maternal age (AMA) [1]. This strategy was based upon an assumption that aneuploidy, which is associate with AMA, is an important factor for implantation failure and therefore such strategy can improve the reproductive outcome [2]. However, a famous randomized study published by Mastenbroek and colleagues reported that such FISH-based PGS with Day 3 biopsy did not confer advantages in women with AMA regarding the live-birth rates [3], and it is then called the “Mastenbroek controversy”. This unexpected finding was subsequently proved by many randomized controlled trials [4] except in one series [5]. Some other researchers thus proposed different strategies including adopting different timing of biopsy (trophectoderm biopsy versus cleavage-stage blastomere biopsy), different molecular technologies which can detect all 24 chromosomes instead of only a few selected chromosomes (which is a major limitation of FISH), and different timing of embryo transfer (fresh or frozen) [1], [6], [7]. Mounting evidences had suggested blastomere biopsy at Day 3 cleavage stage embryos does impair the implantation potential whereas trophectoderm biopsy at Day 5/6 blastocyst does not [8]–[10]. Meanwhile, it is straightforward that using more sophisticated molecular tools may select a better or “more normal” embryo. Competing technologies include array comparative genomic hybridization (CGH), single nucleotide polymorphism (SNP) array, quantitative real-time polymerase chain reaction (qPCR), and next generation sequencing (NGS) [11]. Amongst them, NGS remains to be mostly investigational because of the resources and cost of bioinformatics analyses [12], [13], and therefore the most feasible tools at the present seem to be qPCR and array-based technologies, including SNP array and array CGH [8], [14]–[18]. It is now known that mosaicism is a common phenomenon in early human embryo development, and therefore how to select the embryos with best implantation potential becomes a vital question [19]. It is shown by the research groups who proposed for qPCR that qPCR is superior to array CGH since it enjoys a lower false-positive rate for aneuploidy detection (0 % for qPCR and 5.4 % for array CGH in 122 embryos analyzed), and thereby preserving more embryos available for transfer and ensues a better reproductive outcome [14]. There are now commercially available PGS platforms using array CGH in the market (such as those produced by BlueGnome, UK) and has become very popular because of the feasibility of an easy-to-use system, especially for in vitro fertilization (IVF) centers without a genetic lab as a supporting resort [11]. However, since the outcomes achieved by the team proposing qPCR are compellingly excellent. It thus poses stress upon researchers of the similar field to verify qPCR as a replacement or an alternative to the current popular PGS by array CGH platforms [14]. In this small verification study, we aimed to assess the PGS by qPCR strategy coupled with trophectoderm biopsy at Day 5/6 blastocyst stage. In the initial stage we first verified qPCR only at selected common aneuploidies including chromosome 13, 18, 21, X, and Y (these aneuploidies can be confirmed by FISH). The reasons we assessed only selected chromosomes are three: First, the detail design and experimental conditions regarding qPCR on all 24 chromosomes are patent protected and we need staged efforts to devise and validate our own. Therefore, in this study we adopted a method called locked nucleic acid (LNA) technology to design our qPCR platform [20]; Second, for a rapid overnight diagnosis which is warranted for fresh embryo transfer (FET), it takes a qPCR machine with a 384-well for only one embryo [18], which may not be feasible in real settings in most genetic labs; Third, we believe the “Mastenbroek controversy” may result not only from lacking an effective genotyping system to screen a “better” embryo, but also from the proven impairment of the implantation potential when biopsy at Day 3 cleavage stage embryos [3], [4], [6]. The team proposing qPCR in one hand arguing qPCR is better than FISH because it can examine all 24 chromosomes but in the other hand claiming despite array CGH should have a better resolution than qPCR whereas implying array CGH is “too sensitive” since it has a more false-positive rate when SNP array is used as a gold standard [14], [16], [18]. We thus are interested to know if it is feasible by using a limited qPCR strategy with examination of only selected common aneuploidies (similar to FISH) coupled with trophectoderm biopsy of Day 5/6 blastocyst stage embryos can still achieve a favorable reproductive outcome. It is ethical since it is by theory better than the trial protocol reported in the “Mastenbroek controversy” trial [3], in which a similar number of aneuploidies was assessed by FISH but the biopsy timing was set at Day 3 (when cleavage-stage embryos), a timing now being recognized to be inferior to Day 5/6 blastocyst stage regarding the implantation potential [9]. After the feasibility is confirmed in our setting, it is then justified to expand the repertoire of aneuploidies to 9 chromosomes (13, 15, 16, 17, 18, 21, 22, X and Y. See Rubio et al., 2013 a[5]), or 11 chromosomes (7, 9, 11, 13, 14, 15, 18, 21, 22, X and Y. These are the most commonly found trisomies at abortus. See Wang et al., 2014 [21]) and eventually to 24 chromosomes (1–22, X and Y. See Treff et al., 2012 [1]) in the future. Results Pre-clinical validation Pre-clinical validation for the PGS by qPCR was performed on 54 surplus frozen embryos (Additional file 1: Table S1). Fifty-three of these embryos were successfully diagnosed and the successful diagnostic rate was 98.1 % (53/54). Twenty-four embryos were found to carry aneuploidy on tested chromosomes and the rate of aneuploidy was thus 44.4 % (24/53). The only embryo failed to provide a confirmed diagnosis showed poor qPCR signal, possibly due to a low amount of or degraded DNA in the biopsied sample. FISH diagnosis for that embryo was negative for numerical disorders involving chromosome 13, 18, 21, X, and Y. The remaining cells of this biopsied embryo were sent for array CGH and the diagnosis was trisomy 22. Clinical verification A total of 72 embryos at Day 5/6 blastocyst stage from 13 patients were biopsied and assessed, and all (100 %) of them were successfully diagnosed (Table 1 and Additional file 1: Table S1). Twenty-eight (38.9 %) of these embryos were aneuploid, which included 4 monosomy 13, 5 monosomy 21, 7 trisomy 13, 1 trisomy 18, 9 trisomy 21, 1 monosomy 13 + trisomy 18, and 1 monosomy 18 + monosomy 21. The remaining 44 (61.1 %) embryos were euploidy, of which 28 were transferred into the 13 patients in 13 cycles. The average number of the embryos being transferred is 2.15, actually only 1–2 embryos were transferred in all women of the Group B (non-AMA group, n = 10). The only 2 women who were transferred for 4 embryos are both in the Group A (AMA group, n = 3). The transfer procedure follows the medical rule of Taiwan that the upper limit of the number of the embryos being transferred is 4. These transfers achieved 8 pregnancies, and therefore the clinical pregnancy rate was 61.5 % (8/13) per cycle. However, one early abortion was noted and the ongoing pregnancy rate up to the second trimester was 53.8 % (7/13) per transfer cycle. There were no monozygotic twinning and a total of 11 sacs were noted. Three women who were transferred for 2 embryos in the Group B had singleton pregnancies, and two women who were transferred for 2 embryos in the Group B and 2 women who were transferred for 4 embryos in the Group A had dizygotic twin pregnancies (Additional file 1: Table S1). The sustained implantation rate was thus 39.3 % (11/28) per transferred fresh embryo (Table 1). Totally, there are 2 of 3 women in Group A had ongoing pregnancies. Six women in Group B had clinical pregnancies, and 5 of them had sustained ongoing pregnancies. There are no statistical significant differences noted between these two groups regarding aneuploidy (Chi square test, p = 0.094), clinical pregnancy (Fisher's exact test, p = 1), and ongoing pregnancy (Fisher's exact test, p = 1). Array CGH for the only abortion case revealed the karyotype was 46,XY. Table 1. Clinical outcomes of PGS by qPCR with fresh embryo transfer (FET) verification series Discussion The major difficulty of comprehensive chromosome screening (CCS) by qPCR for PGS is to devise a primer set to successfully amplify the molecular markers being selected in a single experiment by optimizing the conditions for melting temperatures and experimental time intervals. In this study we successfully adopted a smart design called LNA methodology [20], [22]–[24] for qPCR PGS and verified the strategy by achieving a favorable ongoing pregnancy rate of 53.8 %. We first validated the genotyping platform we devised by achieving an almost 100 % successful diagnosis rate (98.1 %), and then verified the whole protocol of qPCR PGS followed by FET. Since the sample size is small (n = 13 cycles), it is not surprising that we failed to observe significant difference regarding the aneuploidy rates between the AMA (n = 3) and the non-AMA group (n = 10) in our patients. Meanwhile, the aneuploidy rate in our series (38.9 %) is apparently much lower than the rates observed by 24-chromosome qPCR or array CGH (more than 60 %), which is obviously due to the fact that we only selected a limited set of common aneuploidies that can ensue live births in humans [14], [15]. It has been a routine practice in our settings that PGS by FISH at Day 3 cleavage-stage embryos since 2005 and it is ethical for us to offer our patients who opted for this alternative protocol a strategy which has an advantage of an established less detrimental effect upon implantation potential by changing the timing of biopsy from Day 3 to Day 5/6 and the same repertoire of common aneuploidies being tested (but simply using qPCR instead of FISH) to enable FET[9]. The reason only trisomy 13, 18, 21, and numerical disorders involving X and Y were tested in our settings is because these are the only aneuploidies that may result in live births in humans. For people who opted for this strategy and came to our clinics, their major concern is more focused at aneuploidies instead of live birth rates. In addition, it is well recognized that despite fewer embryos would be available for transfer if being cultured in vitro onto the blastocyst stage when compared with the cleavage-stage embryos, the disadvantage may be offset by the fact that the aneuploidy rate of the blastocysts is much lower than of the cleavage-stage embryos. An observational study had reported that karyotypic evolution did exist in early human embryogenesis, in which some cleavage stage embryos classified as aneuploidy by FISH eventually developed into euploid blastocysts if by SNP array[25]. A recent randomized study even pointed out that FISH-based PGS conferred advantages at women who are in the AMA group, but not in those with repeated implantation failures group, as long as the biopsy was performed at the blastocyst stage [5]. A recent randomized trial (the BEST trial) had demonstrated that qPCR PGS CCS followed by single embryo transfer (SET) can enhance the feasibility of SET by selecting a single euploid embryo with high reproductive potential without compromising the delivery rates when compared with transfer of two unselected embryos, which instead resulted in more multiple pregnancies and the associated complications such as preterm delivery, low birth weight, and NICU (Neonatal Intensive Care Unit) admission [26]. We admit our pilot verification study had suffered from the fact that only common aneuploidies of five chromosome pairs (13, 18, 21, X, and Y) were screened and apparently it is the reason that the implantation rate per embryo in our study (39.3 %) is much lower than that in the BEST trial (69 %) [26]. However, our patients were all informed consented and knew the disadvantage before joining the study, and the limitations of the local setting were clearly disclosed to the patients. The patients were free to choose another PGS protocol by array CGH (Option 2, please refer to Patients and methods section) instead. Conclusions The merit of our work is that this is the first effort from research groups other than the original group who proposed the qPCR CCS (Reproductive Medicine Associates of New Jersey, Morristown, NJ, USA) trying to validate and verify this novel genotyping platform which was developed in-house by ourselves. Despite our results did not have matched controls such as those cycles receiving IVF but without PGS, the ongoing pregnancy rate is similar to a previous PGD series from our setting (53.8 % versus 50 %, please refer to Chang et al., 2013 [27]), indicating this protocol is clinically feasible. Future efforts will be made upon expanding the repertoire of the PGS by qPCR CCS to all 24 chromosomes, as well as carefully-designed randomized controlled trials to compare this genotyping platform with others (including no-screening, array CGH, and NGS). Patients and methods Design and selection of molecular markers for qPCR PGS An in-house screening system by dual-color qPCR was developed with specific primers for the targeted loci (situated at chromosome 13, 18, 21, X, and Y) and one internal control locus (situated at chromosome 1, the reason we chose chromosome 1 as control is because it is the least found trisomy in humans, see Wang et al., 2014 [21]). A total of 16 targeted loci and one control locus were selected. For the detailed information about the targeted loci, please refer to Table 2. Specific primers for amplification of each locus were designed by using the software Oligo 6.71 (Molecular Biology Insights, Colorado, USA), and all primer sets are flanked on each side of the exon-intron boundary to avoid possible mRNA interference. The applications of LNA-modified probes are well-established for quantifying levels of gene expression with an advantage of reducing complexity [22], [24], but has not been applied to the detection of chromosomal copy number alterations. Two kinds of LNA-modified probes, labeled with FAM™ and HEX™ (Integrated DNA Technologies, Iowa, USA) at the 5’-end, were used for quantifying the genomic copy numbers of targeted loci and control. Table 2. Summary of the targeted regions for screening of common aneuploidies Validation of qPCR in detecting common aneuploidies Each 5 cells separated from the cell lines of known common aneuploidies (including trisomy 21, trisomy 13, trisomy 18, 47,XXY, and 45,X) were processed by cell lysis of proteinase K, and the products were subjected to a 50-μL reaction volume of multiplex nested-PCR amplication for 18 cycles using an Applied Biosystems Veriti thermal cycler (Life Technologies, California, USA), and then the PCR products were purified using Agencourt AMPure XP system (Beckman Coulter, California, USA). Dual color hydrolysis probe assays were performed in triplicate to normalize and simpify calculation and to evaluate chromosomal copies, using Lightcycler 480 probes Master (Roche, Mannheim, Germany), a 20-μL reaction volume, a 96-well plate, and a 7 Light Cycler 480 Real-Time PCR System, as recommended by the supplier (Roche, Mannheim, Germany). Each well contains a particular target, and a common control reaction. A unique method of the standard delta delta threshold cycle (ΔΔCp) method was used for relative quantification. In our experiments, the Cp variation of all HEX™ reactions obtained for each well of the same sample will be controlled and ranged in less than 0.2, indicating the test sample was evenly distributed to each well. Each chromosome-specific ΔCp was calculated from the Cp of the FAM™ reactions targeting a specific chromosome minus the control Cp of the HEX™ reactions targeting the chromosome 1 within the same well. The same process was applied to individually determine the ΔCp for each targeted chromosome of the test sample, including reference set of normal male cell lines [BCRC number: 08C0011, 08C0012, 08C0013, 08C0021 and 08C0025]. Each chromosome- specific ΔCp was then normalized to the average chromosome-specific ΔCp values derived from the same evaluation of the reference set, which had been confirmed by FISH method. The calibrated chromosome-specific ΔCp values were used to calculate fold change by considering the ΔΔCp values as the negative exponent of 2, as previously described [18], [23]. The methodology was designed to specifically identify whole-chromosome but not segmental aneuploidy. The flowchart and diagram of the in-house qPCR PGS system were illustrated in Fig. 1. This qPCR was capable of accurate aneuploidy screening in 4 h, which allowed rapid evaluation of the trophectoderm biopsies and therefore provided a feasible opportunity for subsequent FET. thumbnailFig. 1. The diagram of the in-house qPCR PGS system. The flowchart of (a) detection of common aneuploidies, and (b) signal normalization and data analysis Pre-clinical validation upon surplus frozen embryos Fifty-four thawed frozen embryos at blastocyst stage were biopsied and sent for qPCR and FISH analyses. These embryos were retrieved from the surplus frozen embryos of couples who already conceived and would be discarded if not being investigated for research purpose. In those embryos diagnosed with common chromosome aneuploidies, array CGH was used to confirm the diagnoses. Clinical verification for fresh embryos In our setting we used to offer two PGS protocols. One option (Option 1) is Day 3 biopsy followed by PGS with FISH and FET; the other option (Option 2) is Day 5/6 trophectoderm biopsy followed by PGS with array CGH and frozen embryo transfer (because array CGH takes time and FET was not feasible at the study period). Only patients who had history of failed IVF (without PGS) for at least once and who opted for Option 1 were given the chance of joining this study as an alternative. All patients chose to join this study were informed consented and their autonomy was fully respected. They could choose to withdraw from the study at any time during the study period and were fully aware of the alternatives, including sticking to Option 1 or instead chose Option 2 without joining the study. During July to October of 2014, 13 infertile couples were enrolled. Among these 13 patients, 3 of them had AMA (37, 43, 45 years old respectively). The mean age of the total 13 patients was 34.1 years. No confounding factors that may affect implantation such as immune aberrations (antiphospholipid antibody syndrome in the mother), balanced translocation carriers (in both couples), and thrombophilias (protein C/S/antithrombin III deficiency) existed in these couples when they were enrolled. Clinical pregnancy was defined as positive urine HCG. Ongoing pregnancy rate (per cycle) was defined as those pregnancies proceeding into second trimester (and for each embryo being transferred, sustained implantation rate was used). 13 couples were classified as those whose age is older or equal to 35 years (Group A), and those whose age is less than 35 years (Group B). Chi square test or Fisher’s exact test was used to compare the reproductive outcomes between the groups regarding the rate of aneuploidy, clinical pregnancy, and ongoing pregnancy. Notably according to the regulations of Taiwan government, the patients would not know the results of the fetal sex. Even in aneuploidies involving sex chromosomes (specifically 47,XXY and 45,X), the results would not be disclosed to patients and only “aneuploidy” would be told to the patients and the aneuploid embryos would not be selected for transfer. In those embryos diagnosed to have chromosomal aneuploidies by qPCR, array CGH was used to confirm the diagnoses. Abbreviations AMA: Advanced maternal age CCS: Comprehensive chromosome screening CGH: Comparative genomic hybridization FET: Fresh embryo transfer FISH: Fluorescence in situ hybridization IVF: in vitro fertilization LNA: Locked nucleic acid NGS: Next generation sequencing PGS: Preimplantation genetic screening qPCR: Quantitative real-time polymerase chain reaction SET: Single embryo transfer SNP: Single nucleotide polymorphism Competing interests The authors declare that they have no competing interests. Authors’ contributions YSY, HFC, SPC, FPT, MC designed the study. SPC, GCM, MC developed the novel in-house genotyping platform. SPC, GCM, WHL, CFL, MC did the experiments. YSY, HFC, FPT, CHW, HDT, MC recruited the patients. YSY, HFC, GCM, THL, MC analyzed the results. YSY, SPC, GCM, WHL, MC wrote the paper. All authors read and approved the final manuscript. Additional file Additional file 1: Table S1.. Summary of the pre-clinical validation (for 54 surplus frozen embryos) and clinical verification (for 13 patients with 54 embryos) of PGS by qPCR. Format: DOCX Size: 32KB Download fileOpen Data Acknowledgements This study is partly funded by a grant from Changhua Christian Hospital, Taiwan to Ming Chen (103-CCH-IST-006). The authors are grateful for the genuine support and advice from Dr. Shou-Jen Kuo, the Superintendent of Changhua Christian Hospital, who also participated in the development of the novel in-house qPCR CCS system. References Treff NR, Scott RT. Methods for comprehensive chromosome screening of oocytes and embryos: capabilities, limitations, and evidence of validity. 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Preimplantation genetic screening using fluorescence in situ hybridization in patients with repetitive implantation failure and advanced maternal age: two randomized trials. Fertil Steril. 2013;99(5):1400–7.a Braude P. Selecting the 'best' embryos: prospects for improvement. Reprod Biomed Online. 2013; 27(6):244-53.Publisher Full Text OpenURL Mastenbroek S, Repping S. Preimplantation genetic screening: back to the future. Hum Reprod. 2014; 29(9):1846-50. PubMed Abstract |Publisher Full Text OpenURL Scott RT, Ferry K, Su J, Tao X, Scott K, Treff NR. Comprehensive chromosome screening is highly predictive of the reproductive potential of human embryos: a prospective, blinded, nonselection study. Fertil Steril. 2012; 97(4):870-5. PubMed Abstract |Publisher Full Text OpenURL Scott KL, Hong KH, Scott RT. Selecting the optimal time to perform biopsy for preimplantation genetic testing. Fertil Steril. 2013;100(3):608-14. 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PubMed Abstract | Publisher Full Text OpenURL Treff NR, Levy B, Su J, Northrop LE, Tao X, Scott RT. SNP microarray-based 24 chromosome aneuploidy screening is significantly more consistent than FISH. Mol Hum Reprod. 2010; 16(8):583-9.PubMed Abstract | Publisher Full Text OpenURL Treff NR, Tao X, Ferry KM, Su J, Taylor D, Scott RT. Development and validation of an accurate quantitative real-time polymerase chain reaction-based assay for human blastocyst comprehensive chromosomal aneuploidy screening. Fertil Steril. 2012; 97(4):819-24. PubMed Abstract | Publisher Full Text OpenURL Vanneste E, Voet T, Le Caignec C, Ampe M, Konings P, Melotte C et al..Chromosome instability is common in human cleavage-stage embryos. Nat Med. 2009; 15(5):577-83. PubMed Abstract |Publisher Full Text OpenURL Brunet E, Corgnali M, Cannata F, Perrouault L, Giovannangeli C.Targeting chromosomal sites with locked nucleic acid-modified triplex-forming oligonucleotides: study of efficiency dependence on DNA nuclear environment. Nucleic Acids Res. 2006; 34(16):4546-53. PubMed Abstract | Publisher Full Text OpenURL Wang BT, Chong TP, Boyar FZ, Kopita KA, Ross LP, El-Naggar MM et al..Abnormalities in spontaneous abortions detected by G-banding and chromosomal microarray analysis (CMA) at a national reference laboratory. Mol Cytogenet. 2014; 22:7-33. OpenURL Mouritzen P, Nielsen PS, Jacobsen N, Noerholmn M, Lomholt C, Pfundheller HM et al.. The ProbeLibrary—expression profiling 99 % of all human genes usingonly 90 dual-labeled real-time PCR probes. Biotechniques. 2004; 37:492-495. OpenURL Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative CT method. 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• 2015: 發表的 《一條龍試管嬰兒qPCR》論文 被Molecular cytogenetics 是一個國際期刊拿來廣告對期刊有好的影響 在這封信裡 發表的論文高居第一名被下載的第一名 發表的qPCR論文 被Molecular cytogenetics 是一個國際 期刊拿來廣告對期刊有好的影響 在這封信裡 發表的論文高居第一名被下載的第一名 七月 發表的論文 被期刊拿來廣告 是最常被下載的論文之一 對期刊有好的影響 在這封信裡 發表的論文高居第一名 http://www.molecular cytogenetics.org/content/8/1/49 http://www.molecularcytogenetics.org/content/8/1/49 內置圖片 1內置圖片 2 內置圖片 4 內置圖片 1內置圖片 3內置圖片 2 ​ BioMed Central news@info.biomedcentral.com 透過 bounce.news.springer.com 21:37 (9 小時前) 寄給 我 Dear Colleague, Below is a selection of the latest, highly accessed articles from the open access journal, Molecular Cytogenetics: - Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification - Partial tetrasomy of the proximal long arm of chromosome 15 in two patients: the significance of the gene dosage in terms of phenotype - Genes on B chromosomes of vertebrates Interested in publishing with BioMed Central? 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Research Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification Yu-Shih Yang1†, Shun-Ping Chang2†, Hsin-Fu Chen13†, Gwo-Chin Ma24†, Wen-Hsiang Lin2, Chi-Fang Lin1, Feng-Po Tsai5, Cheng-Hsuan Wu6, Horng-Der Tsai6, Tsung-Hsien Lee17 and Ming Chen1268* *Corresponding author: Ming Chenmchen_cch@yahoo.com; mingchenmd@gmail.com † Equal contributors Author Affiliations 1 Department of Obstetrics and Gynecology, College of Medicine, National Taiwan University, Taipei, Taiwan 2 Department of Genomic Medicine, and Center for Medical Genetics, Changhua Christian Hospital, Changhua, Taiwan 3 Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei, Taiwan 4 Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, Taiwan 5 Poyuan Women Clinic, Changhua, Taiwan 6 Department of Obstetrics and Gynecology, Changhua Christian Hospital, Changhua, Taiwan 7 Department of Obstetrics and Gynecology, Chung-Shan Medical University, Taichung, Taiwan 8 Department of Life Sciences, Tunghai University, Taichung, Taiwan For all author emails, please log on. Molecular Cytogenetics 2015, 8:49 doi:10.1186/s13039-015-0140-9 Yu-Shih Yang, Shun-Ping Chang, Hsin-Fu Chen and Gwo-Chin Ma contributed equally to this work. The electronic version of this article is the complete one and can be found online at: http://www.molecularcytogenetics.org/content/8/1/49 Received: 11 February 2015 Accepted: 7 May 2015 Published: 8 July 2015 © 2015 Yang et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Abstract Background Aneuploidy is an important etiology of implantation failure and quantitative real-time polymerase chain reaction (qPCR) seems a promising preimplantation genetic screening (PGS) technology to detect aneuploidies. This verification study aimed at verifying the impact on reproductive outcomes in in vitrofertilization (IVF) cycles using fresh embryo transfer (FET) in which the embryos were selected by blastocyst biopsy with qPCR-based PGS in our settings. Results A total of 13 infertile couples with more than once failed in vitro fertilization were enrolled during July to October of 2014. PGS was conducted by qPCR with selectively amplified markers to detect common aneuploidies (chromosomes 13, 18, 21, X, and Y). The design of the qPCR molecular markers adopted the locked nucleic acid (LNA) strategy. The blastocyst biopsy was performed on Day 5/6 and the PGS was done on the same day, which enabled FET. A total of 72 blastocysts were biopsied. Successful diagnoses were established in all embryos and the rate of successful diagnosis was 100 %. The aneuploidy rate was 38.9 % (28/72). 28 embryos were transferred. The clinical pregnancy rate was 61.5 % (8/13) per cycle. Early first trimester abortion was encountered in 1 and the ongoing pregnancy rate was 53.8 % (7/13) per cycle. Conclusion This study verified the favorable outcome of adopting PGS with qPCR + FET in our own setting. Expanding the repertoire of aneuploidies being investigated (from a limited set to all 24 chromosomes) is underway and a randomized study by comparing qPCR and other PGS technologies is warranted. Keywords: Aneuploidy; Blastocyst; Fresh embryo transfer; PGS; qPCR http://www.molecularcytogenetics.org/content/8/1/49 Research Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification Yu-Shih Yang1†, Shun-Ping Chang2†, Hsin-Fu Chen13†, Gwo-Chin Ma24†, Wen-Hsiang Lin2, Chi-Fang Lin1, Feng-Po Tsai5, Cheng-Hsuan Wu6, Horng-Der Tsai6, Tsung-Hsien Lee17 and Ming Chen1268* *Corresponding author: Ming Chenmchen_cch@yahoo.com; mingchenmd@gmail.com † Equal contributors Author Affiliations 1 Department of Obstetrics and Gynecology, College of Medicine, National Taiwan University, Taipei, Taiwan 2 Department of Genomic Medicine, and Center for Medical Genetics, Changhua Christian Hospital, Changhua, Taiwan 3 Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei, Taiwan 4 Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, Taiwan 5 Poyuan Women Clinic, Changhua, Taiwan 6 Department of Obstetrics and Gynecology, Changhua Christian Hospital, Changhua, Taiwan 7 Department of Obstetrics and Gynecology, Chung-Shan Medical University, Taichung, Taiwan 8 Department of Life Sciences, Tunghai University, Taichung, Taiwan For all author emails, please log on. Molecular Cytogenetics 2015, 8:49 doi:10.1186/s13039-015-0140-9 Yu-Shih Yang, Shun-Ping Chang, Hsin-Fu Chen and Gwo-Chin Ma contributed equally to this work. The electronic version of this article is the complete one and can be found online at: http://www.molecularcytogenetics.org/content/8/1/49 Received: 11 February 2015 Accepted: 7 May 2015 Published: 8 July 2015 © 2015 Yang et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Abstract Background Aneuploidy is an important etiology of implantation failure and quantitative real-time polymerase chain reaction (qPCR) seems a promising preimplantation genetic screening (PGS) technology to detect aneuploidies. This verification study aimed at verifying the impact on reproductive outcomes in in vitrofertilization (IVF) cycles using fresh embryo transfer (FET) in which the embryos were selected by blastocyst biopsy with qPCR-based PGS in our settings. Results A total of 13 infertile couples with more than once failed in vitro fertilization were enrolled during July to October of 2014. PGS was conducted by qPCR with selectively amplified markers to detect common aneuploidies (chromosomes 13, 18, 21, X, and Y). The design of the qPCR molecular markers adopted the locked nucleic acid (LNA) strategy. The blastocyst biopsy was performed on Day 5/6 and the PGS was done on the same day, which enabled FET. A total of 72 blastocysts were biopsied. Successful diagnoses were established in all embryos and the rate of successful diagnosis was 100 %. The aneuploidy rate was 38.9 % (28/72). 28 embryos were transferred. The clinical pregnancy rate was 61.5 % (8/13) per cycle. Early first trimester abortion was encountered in 1 and the ongoing pregnancy rate was 53.8 % (7/13) per cycle. Conclusion This study verified the favorable outcome of adopting PGS with qPCR + FET in our own setting. Expanding the repertoire of aneuploidies being investigated (from a limited set to all 24 chromosomes) is underway and a randomized study by comparing qPCR and other PGS technologies is warranted. Keywords: Aneuploidy; Blastocyst; Fresh embryo transfer; PGS; qPCR Background Preimplantation genetic screening (PGS) by Day 3 cleavage stage embryo biopsy followed by examination with fluorescence in situ hybridization (FISH) was once popular strategy for women with advanced maternal age (AMA) [1]. This strategy was based upon an assumption that aneuploidy, which is associate with AMA, is an important factor for implantation failure and therefore such strategy can improve the reproductive outcome [2]. However, a famous randomized study published by Mastenbroek and colleagues reported that such FISH-based PGS with Day 3 biopsy did not confer advantages in women with AMA regarding the live-birth rates [3], and it is then called the “Mastenbroek controversy”. This unexpected finding was subsequently proved by many randomized controlled trials [4] except in one series [5]. Some other researchers thus proposed different strategies including adopting different timing of biopsy (trophectoderm biopsy versus cleavage-stage blastomere biopsy), different molecular technologies which can detect all 24 chromosomes instead of only a few selected chromosomes (which is a major limitation of FISH), and different timing of embryo transfer (fresh or frozen) [1], [6], [7]. Mounting evidences had suggested blastomere biopsy at Day 3 cleavage stage embryos does impair the implantation potential whereas trophectoderm biopsy at Day 5/6 blastocyst does not [8]–[10]. Meanwhile, it is straightforward that using more sophisticated molecular tools may select a better or “more normal” embryo. Competing technologies include array comparative genomic hybridization (CGH), single nucleotide polymorphism (SNP) array, quantitative real-time polymerase chain reaction (qPCR), and next generation sequencing (NGS) [11]. Amongst them, NGS remains to be mostly investigational because of the resources and cost of bioinformatics analyses [12], [13], and therefore the most feasible tools at the present seem to be qPCR and array-based technologies, including SNP array and array CGH [8], [14]–[18]. It is now known that mosaicism is a common phenomenon in early human embryo development, and therefore how to select the embryos with best implantation potential becomes a vital question [19]. It is shown by the research groups who proposed for qPCR that qPCR is superior to array CGH since it enjoys a lower false-positive rate for aneuploidy detection (0 % for qPCR and 5.4 % for array CGH in 122 embryos analyzed), and thereby preserving more embryos available for transfer and ensues a better reproductive outcome [14]. There are now commercially available PGS platforms using array CGH in the market (such as those produced by BlueGnome, UK) and has become very popular because of the feasibility of an easy-to-use system, especially for in vitro fertilization (IVF) centers without a genetic lab as a supporting resort [11]. However, since the outcomes achieved by the team proposing qPCR are compellingly excellent. It thus poses stress upon researchers of the similar field to verify qPCR as a replacement or an alternative to the current popular PGS by array CGH platforms [14]. In this small verification study, we aimed to assess the PGS by qPCR strategy coupled with trophectoderm biopsy at Day 5/6 blastocyst stage. In the initial stage we first verified qPCR only at selected common aneuploidies including chromosome 13, 18, 21, X, and Y (these aneuploidies can be confirmed by FISH). The reasons we assessed only selected chromosomes are three: First, the detail design and experimental conditions regarding qPCR on all 24 chromosomes are patent protected and we need staged efforts to devise and validate our own. Therefore, in this study we adopted a method called locked nucleic acid (LNA) technology to design our qPCR platform [20]; Second, for a rapid overnight diagnosis which is warranted for fresh embryo transfer (FET), it takes a qPCR machine with a 384-well for only one embryo [18], which may not be feasible in real settings in most genetic labs; Third, we believe the “Mastenbroek controversy” may result not only from lacking an effective genotyping system to screen a “better” embryo, but also from the proven impairment of the implantation potential when biopsy at Day 3 cleavage stage embryos [3], [4], [6]. The team proposing qPCR in one hand arguing qPCR is better than FISH because it can examine all 24 chromosomes but in the other hand claiming despite array CGH should have a better resolution than qPCR whereas implying array CGH is “too sensitive” since it has a more false-positive rate when SNP array is used as a gold standard [14], [16], [18]. We thus are interested to know if it is feasible by using a limited qPCR strategy with examination of only selected common aneuploidies (similar to FISH) coupled with trophectoderm biopsy of Day 5/6 blastocyst stage embryos can still achieve a favorable reproductive outcome. It is ethical since it is by theory better than the trial protocol reported in the “Mastenbroek controversy” trial [3], in which a similar number of aneuploidies was assessed by FISH but the biopsy timing was set at Day 3 (when cleavage-stage embryos), a timing now being recognized to be inferior to Day 5/6 blastocyst stage regarding the implantation potential [9]. After the feasibility is confirmed in our setting, it is then justified to expand the repertoire of aneuploidies to 9 chromosomes (13, 15, 16, 17, 18, 21, 22, X and Y. See Rubio et al., 2013 a[5]), or 11 chromosomes (7, 9, 11, 13, 14, 15, 18, 21, 22, X and Y. These are the most commonly found trisomies at abortus. See Wang et al., 2014 [21]) and eventually to 24 chromosomes (1–22, X and Y. See Treff et al., 2012 [1]) in the future. Results Pre-clinical validation Pre-clinical validation for the PGS by qPCR was performed on 54 surplus frozen embryos (Additional file 1: Table S1). Fifty-three of these embryos were successfully diagnosed and the successful diagnostic rate was 98.1 % (53/54). Twenty-four embryos were found to carry aneuploidy on tested chromosomes and the rate of aneuploidy was thus 44.4 % (24/53). The only embryo failed to provide a confirmed diagnosis showed poor qPCR signal, possibly due to a low amount of or degraded DNA in the biopsied sample. FISH diagnosis for that embryo was negative for numerical disorders involving chromosome 13, 18, 21, X, and Y. The remaining cells of this biopsied embryo were sent for array CGH and the diagnosis was trisomy 22. Clinical verification A total of 72 embryos at Day 5/6 blastocyst stage from 13 patients were biopsied and assessed, and all (100 %) of them were successfully diagnosed (Table 1 and Additional file 1: Table S1). Twenty-eight (38.9 %) of these embryos were aneuploid, which included 4 monosomy 13, 5 monosomy 21, 7 trisomy 13, 1 trisomy 18, 9 trisomy 21, 1 monosomy 13 + trisomy 18, and 1 monosomy 18 + monosomy 21. The remaining 44 (61.1 %) embryos were euploidy, of which 28 were transferred into the 13 patients in 13 cycles. The average number of the embryos being transferred is 2.15, actually only 1–2 embryos were transferred in all women of the Group B (non-AMA group, n = 10). The only 2 women who were transferred for 4 embryos are both in the Group A (AMA group, n = 3). The transfer procedure follows the medical rule of Taiwan that the upper limit of the number of the embryos being transferred is 4. These transfers achieved 8 pregnancies, and therefore the clinical pregnancy rate was 61.5 % (8/13) per cycle. However, one early abortion was noted and the ongoing pregnancy rate up to the second trimester was 53.8 % (7/13) per transfer cycle. There were no monozygotic twinning and a total of 11 sacs were noted. Three women who were transferred for 2 embryos in the Group B had singleton pregnancies, and two women who were transferred for 2 embryos in the Group B and 2 women who were transferred for 4 embryos in the Group A had dizygotic twin pregnancies (Additional file 1: Table S1). The sustained implantation rate was thus 39.3 % (11/28) per transferred fresh embryo (Table 1). Totally, there are 2 of 3 women in Group A had ongoing pregnancies. Six women in Group B had clinical pregnancies, and 5 of them had sustained ongoing pregnancies. There are no statistical significant differences noted between these two groups regarding aneuploidy (Chi square test, p = 0.094), clinical pregnancy (Fisher's exact test, p = 1), and ongoing pregnancy (Fisher's exact test, p = 1). Array CGH for the only abortion case revealed the karyotype was 46,XY. Table 1. Clinical outcomes of PGS by qPCR with fresh embryo transfer (FET) verification series Discussion The major difficulty of comprehensive chromosome screening (CCS) by qPCR for PGS is to devise a primer set to successfully amplify the molecular markers being selected in a single experiment by optimizing the conditions for melting temperatures and experimental time intervals. In this study we successfully adopted a smart design called LNA methodology [20], [22]–[24] for qPCR PGS and verified the strategy by achieving a favorable ongoing pregnancy rate of 53.8 %. We first validated the genotyping platform we devised by achieving an almost 100 % successful diagnosis rate (98.1 %), and then verified the whole protocol of qPCR PGS followed by FET. Since the sample size is small (n = 13 cycles), it is not surprising that we failed to observe significant difference regarding the aneuploidy rates between the AMA (n = 3) and the non-AMA group (n = 10) in our patients. Meanwhile, the aneuploidy rate in our series (38.9 %) is apparently much lower than the rates observed by 24-chromosome qPCR or array CGH (more than 60 %), which is obviously due to the fact that we only selected a limited set of common aneuploidies that can ensue live births in humans [14], [15]. It has been a routine practice in our settings that PGS by FISH at Day 3 cleavage-stage embryos since 2005 and it is ethical for us to offer our patients who opted for this alternative protocol a strategy which has an advantage of an established less detrimental effect upon implantation potential by changing the timing of biopsy from Day 3 to Day 5/6 and the same repertoire of common aneuploidies being tested (but simply using qPCR instead of FISH) to enable FET[9]. The reason only trisomy 13, 18, 21, and numerical disorders involving X and Y were tested in our settings is because these are the only aneuploidies that may result in live births in humans. For people who opted for this strategy and came to our clinics, their major concern is more focused at aneuploidies instead of live birth rates. In addition, it is well recognized that despite fewer embryos would be available for transfer if being cultured in vitro onto the blastocyst stage when compared with the cleavage-stage embryos, the disadvantage may be offset by the fact that the aneuploidy rate of the blastocysts is much lower than of the cleavage-stage embryos. An observational study had reported that karyotypic evolution did exist in early human embryogenesis, in which some cleavage stage embryos classified as aneuploidy by FISH eventually developed into euploid blastocysts if by SNP array[25]. A recent randomized study even pointed out that FISH-based PGS conferred advantages at women who are in the AMA group, but not in those with repeated implantation failures group, as long as the biopsy was performed at the blastocyst stage [5]. A recent randomized trial (the BEST trial) had demonstrated that qPCR PGS CCS followed by single embryo transfer (SET) can enhance the feasibility of SET by selecting a single euploid embryo with high reproductive potential without compromising the delivery rates when compared with transfer of two unselected embryos, which instead resulted in more multiple pregnancies and the associated complications such as preterm delivery, low birth weight, and NICU (Neonatal Intensive Care Unit) admission [26]. We admit our pilot verification study had suffered from the fact that only common aneuploidies of five chromosome pairs (13, 18, 21, X, and Y) were screened and apparently it is the reason that the implantation rate per embryo in our study (39.3 %) is much lower than that in the BEST trial (69 %) [26]. However, our patients were all informed consented and knew the disadvantage before joining the study, and the limitations of the local setting were clearly disclosed to the patients. The patients were free to choose another PGS protocol by array CGH (Option 2, please refer to Patients and methods section) instead. Conclusions The merit of our work is that this is the first effort from research groups other than the original group who proposed the qPCR CCS (Reproductive Medicine Associates of New Jersey, Morristown, NJ, USA) trying to validate and verify this novel genotyping platform which was developed in-house by ourselves. Despite our results did not have matched controls such as those cycles receiving IVF but without PGS, the ongoing pregnancy rate is similar to a previous PGD series from our setting (53.8 % versus 50 %, please refer to Chang et al., 2013 [27]), indicating this protocol is clinically feasible. Future efforts will be made upon expanding the repertoire of the PGS by qPCR CCS to all 24 chromosomes, as well as carefully-designed randomized controlled trials to compare this genotyping platform with others (including no-screening, array CGH, and NGS). Patients and methods Design and selection of molecular markers for qPCR PGS An in-house screening system by dual-color qPCR was developed with specific primers for the targeted loci (situated at chromosome 13, 18, 21, X, and Y) and one internal control locus (situated at chromosome 1, the reason we chose chromosome 1 as control is because it is the least found trisomy in humans, see Wang et al., 2014 [21]). A total of 16 targeted loci and one control locus were selected. For the detailed information about the targeted loci, please refer to Table 2. Specific primers for amplification of each locus were designed by using the software Oligo 6.71 (Molecular Biology Insights, Colorado, USA), and all primer sets are flanked on each side of the exon-intron boundary to avoid possible mRNA interference. The applications of LNA-modified probes are well-established for quantifying levels of gene expression with an advantage of reducing complexity [22], [24], but has not been applied to the detection of chromosomal copy number alterations. Two kinds of LNA-modified probes, labeled with FAM™ and HEX™ (Integrated DNA Technologies, Iowa, USA) at the 5’-end, were used for quantifying the genomic copy numbers of targeted loci and control. Table 2. Summary of the targeted regions for screening of common aneuploidies Validation of qPCR in detecting common aneuploidies Each 5 cells separated from the cell lines of known common aneuploidies (including trisomy 21, trisomy 13, trisomy 18, 47,XXY, and 45,X) were processed by cell lysis of proteinase K, and the products were subjected to a 50-μL reaction volume of multiplex nested-PCR amplication for 18 cycles using an Applied Biosystems Veriti thermal cycler (Life Technologies, California, USA), and then the PCR products were purified using Agencourt AMPure XP system (Beckman Coulter, California, USA). Dual color hydrolysis probe assays were performed in triplicate to normalize and simpify calculation and to evaluate chromosomal copies, using Lightcycler 480 probes Master (Roche, Mannheim, Germany), a 20-μL reaction volume, a 96-well plate, and a 7 Light Cycler 480 Real-Time PCR System, as recommended by the supplier (Roche, Mannheim, Germany). Each well contains a particular target, and a common control reaction. A unique method of the standard delta delta threshold cycle (ΔΔCp) method was used for relative quantification. In our experiments, the Cp variation of all HEX™ reactions obtained for each well of the same sample will be controlled and ranged in less than 0.2, indicating the test sample was evenly distributed to each well. Each chromosome-specific ΔCp was calculated from the Cp of the FAM™ reactions targeting a specific chromosome minus the control Cp of the HEX™ reactions targeting the chromosome 1 within the same well. The same process was applied to individually determine the ΔCp for each targeted chromosome of the test sample, including reference set of normal male cell lines [BCRC number: 08C0011, 08C0012, 08C0013, 08C0021 and 08C0025]. Each chromosome- specific ΔCp was then normalized to the average chromosome-specific ΔCp values derived from the same evaluation of the reference set, which had been confirmed by FISH method. The calibrated chromosome-specific ΔCp values were used to calculate fold change by considering the ΔΔCp values as the negative exponent of 2, as previously described [18], [23]. The methodology was designed to specifically identify whole-chromosome but not segmental aneuploidy. The flowchart and diagram of the in-house qPCR PGS system were illustrated in Fig. 1. This qPCR was capable of accurate aneuploidy screening in 4 h, which allowed rapid evaluation of the trophectoderm biopsies and therefore provided a feasible opportunity for subsequent FET. thumbnailFig. 1. The diagram of the in-house qPCR PGS system. The flowchart of (a) detection of common aneuploidies, and (b) signal normalization and data analysis Pre-clinical validation upon surplus frozen embryos Fifty-four thawed frozen embryos at blastocyst stage were biopsied and sent for qPCR and FISH analyses. These embryos were retrieved from the surplus frozen embryos of couples who already conceived and would be discarded if not being investigated for research purpose. In those embryos diagnosed with common chromosome aneuploidies, array CGH was used to confirm the diagnoses. Clinical verification for fresh embryos In our setting we used to offer two PGS protocols. One option (Option 1) is Day 3 biopsy followed by PGS with FISH and FET; the other option (Option 2) is Day 5/6 trophectoderm biopsy followed by PGS with array CGH and frozen embryo transfer (because array CGH takes time and FET was not feasible at the study period). Only patients who had history of failed IVF (without PGS) for at least once and who opted for Option 1 were given the chance of joining this study as an alternative. All patients chose to join this study were informed consented and their autonomy was fully respected. They could choose to withdraw from the study at any time during the study period and were fully aware of the alternatives, including sticking to Option 1 or instead chose Option 2 without joining the study. During July to October of 2014, 13 infertile couples were enrolled. Among these 13 patients, 3 of them had AMA (37, 43, 45 years old respectively). The mean age of the total 13 patients was 34.1 years. No confounding factors that may affect implantation such as immune aberrations (antiphospholipid antibody syndrome in the mother), balanced translocation carriers (in both couples), and thrombophilias (protein C/S/antithrombin III deficiency) existed in these couples when they were enrolled. Clinical pregnancy was defined as positive urine HCG. Ongoing pregnancy rate (per cycle) was defined as those pregnancies proceeding into second trimester (and for each embryo being transferred, sustained implantation rate was used). 13 couples were classified as those whose age is older or equal to 35 years (Group A), and those whose age is less than 35 years (Group B). Chi square test or Fisher’s exact test was used to compare the reproductive outcomes between the groups regarding the rate of aneuploidy, clinical pregnancy, and ongoing pregnancy. Notably according to the regulations of Taiwan government, the patients would not know the results of the fetal sex. Even in aneuploidies involving sex chromosomes (specifically 47,XXY and 45,X), the results would not be disclosed to patients and only “aneuploidy” would be told to the patients and the aneuploid embryos would not be selected for transfer. In those embryos diagnosed to have chromosomal aneuploidies by qPCR, array CGH was used to confirm the diagnoses. Abbreviations AMA: Advanced maternal age CCS: Comprehensive chromosome screening CGH: Comparative genomic hybridization FET: Fresh embryo transfer FISH: Fluorescence in situ hybridization IVF: in vitro fertilization LNA: Locked nucleic acid NGS: Next generation sequencing PGS: Preimplantation genetic screening qPCR: Quantitative real-time polymerase chain reaction SET: Single embryo transfer SNP: Single nucleotide polymorphism Competing interests The authors declare that they have no competing interests. Authors’ contributions YSY, HFC, SPC, FPT, MC designed the study. SPC, GCM, MC developed the novel in-house genotyping platform. SPC, GCM, WHL, CFL, MC did the experiments. YSY, HFC, FPT, CHW, HDT, MC recruited the patients. YSY, HFC, GCM, THL, MC analyzed the results. YSY, SPC, GCM, WHL, MC wrote the paper. All authors read and approved the final manuscript. Additional file Additional file 1: Table S1.. Summary of the pre-clinical validation (for 54 surplus frozen embryos) and clinical verification (for 13 patients with 54 embryos) of PGS by qPCR. Format: DOCX Size: 32KB Download fileOpen Data Acknowledgements This study is partly funded by a grant from Changhua Christian Hospital, Taiwan to Ming Chen (103-CCH-IST-006). The authors are grateful for the genuine support and advice from Dr. Shou-Jen Kuo, the Superintendent of Changhua Christian Hospital, who also participated in the development of the novel in-house qPCR CCS system. References Treff NR, Scott RT. Methods for comprehensive chromosome screening of oocytes and embryos: capabilities, limitations, and evidence of validity. 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• 2015: 發表的 《一條龍試管嬰兒qPCR》論文 被Molecular cytogenetics 是一個國際期刊拿來廣告對期刊有好的影響 在這封信裡 發表的論文高居第一名被下載的第一名 發表的qPCR論文 被Molecular cytogenetics 是一個國際 期刊拿來廣告對期刊有好的影響 在這封信裡 發表的論文高居第一名被下載的第一名 七月 發表的論文 被期刊拿來廣告 是最常被下載的論文之一 對期刊有好的影響 在這封信裡 發表的論文高居第一名 http://www.molecular cytogenetics.org/content/8/1/49 http://www.molecularcytogenetics.org/content/8/1/49 內置圖片 1內置圖片 2 內置圖片 4 內置圖片 1內置圖片 3內置圖片 2 ​ BioMed Central news@info.biomedcentral.com 透過 bounce.news.springer.com 21:37 (9 小時前) 寄給 我 Dear Colleague, Below is a selection of the latest, highly accessed articles from the open access journal, Molecular Cytogenetics: - Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification - Partial tetrasomy of the proximal long arm of chromosome 15 in two patients: the significance of the gene dosage in terms of phenotype - Genes on B chromosomes of vertebrates Interested in publishing with BioMed Central? 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Research Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification Yu-Shih Yang1†, Shun-Ping Chang2†, Hsin-Fu Chen13†, Gwo-Chin Ma24†, Wen-Hsiang Lin2, Chi-Fang Lin1, Feng-Po Tsai5, Cheng-Hsuan Wu6, Horng-Der Tsai6, Tsung-Hsien Lee17 and Ming Chen1268* *Corresponding author: Ming Chenmchen_cch@yahoo.com; mingchenmd@gmail.com † Equal contributors Author Affiliations 1 Department of Obstetrics and Gynecology, College of Medicine, National Taiwan University, Taipei, Taiwan 2 Department of Genomic Medicine, and Center for Medical Genetics, Changhua Christian Hospital, Changhua, Taiwan 3 Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei, Taiwan 4 Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, Taiwan 5 Poyuan Women Clinic, Changhua, Taiwan 6 Department of Obstetrics and Gynecology, Changhua Christian Hospital, Changhua, Taiwan 7 Department of Obstetrics and Gynecology, Chung-Shan Medical University, Taichung, Taiwan 8 Department of Life Sciences, Tunghai University, Taichung, Taiwan For all author emails, please log on. Molecular Cytogenetics 2015, 8:49 doi:10.1186/s13039-015-0140-9 Yu-Shih Yang, Shun-Ping Chang, Hsin-Fu Chen and Gwo-Chin Ma contributed equally to this work. The electronic version of this article is the complete one and can be found online at: http://www.molecularcytogenetics.org/content/8/1/49 Received: 11 February 2015 Accepted: 7 May 2015 Published: 8 July 2015 © 2015 Yang et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Abstract Background Aneuploidy is an important etiology of implantation failure and quantitative real-time polymerase chain reaction (qPCR) seems a promising preimplantation genetic screening (PGS) technology to detect aneuploidies. This verification study aimed at verifying the impact on reproductive outcomes in in vitrofertilization (IVF) cycles using fresh embryo transfer (FET) in which the embryos were selected by blastocyst biopsy with qPCR-based PGS in our settings. Results A total of 13 infertile couples with more than once failed in vitro fertilization were enrolled during July to October of 2014. PGS was conducted by qPCR with selectively amplified markers to detect common aneuploidies (chromosomes 13, 18, 21, X, and Y). The design of the qPCR molecular markers adopted the locked nucleic acid (LNA) strategy. The blastocyst biopsy was performed on Day 5/6 and the PGS was done on the same day, which enabled FET. A total of 72 blastocysts were biopsied. Successful diagnoses were established in all embryos and the rate of successful diagnosis was 100 %. The aneuploidy rate was 38.9 % (28/72). 28 embryos were transferred. The clinical pregnancy rate was 61.5 % (8/13) per cycle. Early first trimester abortion was encountered in 1 and the ongoing pregnancy rate was 53.8 % (7/13) per cycle. Conclusion This study verified the favorable outcome of adopting PGS with qPCR + FET in our own setting. Expanding the repertoire of aneuploidies being investigated (from a limited set to all 24 chromosomes) is underway and a randomized study by comparing qPCR and other PGS technologies is warranted. Keywords: Aneuploidy; Blastocyst; Fresh embryo transfer; PGS; qPCR http://www.molecularcytogenetics.org/content/8/1/49 Research Preimplantation genetic screening of blastocysts by multiplex qPCR followed by fresh embryo transfer: validation and verification Yu-Shih Yang1†, Shun-Ping Chang2†, Hsin-Fu Chen13†, Gwo-Chin Ma24†, Wen-Hsiang Lin2, Chi-Fang Lin1, Feng-Po Tsai5, Cheng-Hsuan Wu6, Horng-Der Tsai6, Tsung-Hsien Lee17 and Ming Chen1268* *Corresponding author: Ming Chenmchen_cch@yahoo.com; mingchenmd@gmail.com † Equal contributors Author Affiliations 1 Department of Obstetrics and Gynecology, College of Medicine, National Taiwan University, Taipei, Taiwan 2 Department of Genomic Medicine, and Center for Medical Genetics, Changhua Christian Hospital, Changhua, Taiwan 3 Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei, Taiwan 4 Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, Taiwan 5 Poyuan Women Clinic, Changhua, Taiwan 6 Department of Obstetrics and Gynecology, Changhua Christian Hospital, Changhua, Taiwan 7 Department of Obstetrics and Gynecology, Chung-Shan Medical University, Taichung, Taiwan 8 Department of Life Sciences, Tunghai University, Taichung, Taiwan For all author emails, please log on. Molecular Cytogenetics 2015, 8:49 doi:10.1186/s13039-015-0140-9 Yu-Shih Yang, Shun-Ping Chang, Hsin-Fu Chen and Gwo-Chin Ma contributed equally to this work. The electronic version of this article is the complete one and can be found online at: http://www.molecularcytogenetics.org/content/8/1/49 Received: 11 February 2015 Accepted: 7 May 2015 Published: 8 July 2015 © 2015 Yang et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Abstract Background Aneuploidy is an important etiology of implantation failure and quantitative real-time polymerase chain reaction (qPCR) seems a promising preimplantation genetic screening (PGS) technology to detect aneuploidies. This verification study aimed at verifying the impact on reproductive outcomes in in vitrofertilization (IVF) cycles using fresh embryo transfer (FET) in which the embryos were selected by blastocyst biopsy with qPCR-based PGS in our settings. Results A total of 13 infertile couples with more than once failed in vitro fertilization were enrolled during July to October of 2014. PGS was conducted by qPCR with selectively amplified markers to detect common aneuploidies (chromosomes 13, 18, 21, X, and Y). The design of the qPCR molecular markers adopted the locked nucleic acid (LNA) strategy. The blastocyst biopsy was performed on Day 5/6 and the PGS was done on the same day, which enabled FET. A total of 72 blastocysts were biopsied. Successful diagnoses were established in all embryos and the rate of successful diagnosis was 100 %. The aneuploidy rate was 38.9 % (28/72). 28 embryos were transferred. The clinical pregnancy rate was 61.5 % (8/13) per cycle. Early first trimester abortion was encountered in 1 and the ongoing pregnancy rate was 53.8 % (7/13) per cycle. Conclusion This study verified the favorable outcome of adopting PGS with qPCR + FET in our own setting. Expanding the repertoire of aneuploidies being investigated (from a limited set to all 24 chromosomes) is underway and a randomized study by comparing qPCR and other PGS technologies is warranted. Keywords: Aneuploidy; Blastocyst; Fresh embryo transfer; PGS; qPCR Background Preimplantation genetic screening (PGS) by Day 3 cleavage stage embryo biopsy followed by examination with fluorescence in situ hybridization (FISH) was once popular strategy for women with advanced maternal age (AMA) [1]. This strategy was based upon an assumption that aneuploidy, which is associate with AMA, is an important factor for implantation failure and therefore such strategy can improve the reproductive outcome [2]. However, a famous randomized study published by Mastenbroek and colleagues reported that such FISH-based PGS with Day 3 biopsy did not confer advantages in women with AMA regarding the live-birth rates [3], and it is then called the “Mastenbroek controversy”. This unexpected finding was subsequently proved by many randomized controlled trials [4] except in one series [5]. Some other researchers thus proposed different strategies including adopting different timing of biopsy (trophectoderm biopsy versus cleavage-stage blastomere biopsy), different molecular technologies which can detect all 24 chromosomes instead of only a few selected chromosomes (which is a major limitation of FISH), and different timing of embryo transfer (fresh or frozen) [1], [6], [7]. Mounting evidences had suggested blastomere biopsy at Day 3 cleavage stage embryos does impair the implantation potential whereas trophectoderm biopsy at Day 5/6 blastocyst does not [8]–[10]. Meanwhile, it is straightforward that using more sophisticated molecular tools may select a better or “more normal” embryo. Competing technologies include array comparative genomic hybridization (CGH), single nucleotide polymorphism (SNP) array, quantitative real-time polymerase chain reaction (qPCR), and next generation sequencing (NGS) [11]. Amongst them, NGS remains to be mostly investigational because of the resources and cost of bioinformatics analyses [12], [13], and therefore the most feasible tools at the present seem to be qPCR and array-based technologies, including SNP array and array CGH [8], [14]–[18]. It is now known that mosaicism is a common phenomenon in early human embryo development, and therefore how to select the embryos with best implantation potential becomes a vital question [19]. It is shown by the research groups who proposed for qPCR that qPCR is superior to array CGH since it enjoys a lower false-positive rate for aneuploidy detection (0 % for qPCR and 5.4 % for array CGH in 122 embryos analyzed), and thereby preserving more embryos available for transfer and ensues a better reproductive outcome [14]. There are now commercially available PGS platforms using array CGH in the market (such as those produced by BlueGnome, UK) and has become very popular because of the feasibility of an easy-to-use system, especially for in vitro fertilization (IVF) centers without a genetic lab as a supporting resort [11]. However, since the outcomes achieved by the team proposing qPCR are compellingly excellent. It thus poses stress upon researchers of the similar field to verify qPCR as a replacement or an alternative to the current popular PGS by array CGH platforms [14]. In this small verification study, we aimed to assess the PGS by qPCR strategy coupled with trophectoderm biopsy at Day 5/6 blastocyst stage. In the initial stage we first verified qPCR only at selected common aneuploidies including chromosome 13, 18, 21, X, and Y (these aneuploidies can be confirmed by FISH). The reasons we assessed only selected chromosomes are three: First, the detail design and experimental conditions regarding qPCR on all 24 chromosomes are patent protected and we need staged efforts to devise and validate our own. Therefore, in this study we adopted a method called locked nucleic acid (LNA) technology to design our qPCR platform [20]; Second, for a rapid overnight diagnosis which is warranted for fresh embryo transfer (FET), it takes a qPCR machine with a 384-well for only one embryo [18], which may not be feasible in real settings in most genetic labs; Third, we believe the “Mastenbroek controversy” may result not only from lacking an effective genotyping system to screen a “better” embryo, but also from the proven impairment of the implantation potential when biopsy at Day 3 cleavage stage embryos [3], [4], [6]. The team proposing qPCR in one hand arguing qPCR is better than FISH because it can examine all 24 chromosomes but in the other hand claiming despite array CGH should have a better resolution than qPCR whereas implying array CGH is “too sensitive” since it has a more false-positive rate when SNP array is used as a gold standard [14], [16], [18]. We thus are interested to know if it is feasible by using a limited qPCR strategy with examination of only selected common aneuploidies (similar to FISH) coupled with trophectoderm biopsy of Day 5/6 blastocyst stage embryos can still achieve a favorable reproductive outcome. It is ethical since it is by theory better than the trial protocol reported in the “Mastenbroek controversy” trial [3], in which a similar number of aneuploidies was assessed by FISH but the biopsy timing was set at Day 3 (when cleavage-stage embryos), a timing now being recognized to be inferior to Day 5/6 blastocyst stage regarding the implantation potential [9]. After the feasibility is confirmed in our setting, it is then justified to expand the repertoire of aneuploidies to 9 chromosomes (13, 15, 16, 17, 18, 21, 22, X and Y. See Rubio et al., 2013 a[5]), or 11 chromosomes (7, 9, 11, 13, 14, 15, 18, 21, 22, X and Y. These are the most commonly found trisomies at abortus. See Wang et al., 2014 [21]) and eventually to 24 chromosomes (1–22, X and Y. See Treff et al., 2012 [1]) in the future. Results Pre-clinical validation Pre-clinical validation for the PGS by qPCR was performed on 54 surplus frozen embryos (Additional file 1: Table S1). Fifty-three of these embryos were successfully diagnosed and the successful diagnostic rate was 98.1 % (53/54). Twenty-four embryos were found to carry aneuploidy on tested chromosomes and the rate of aneuploidy was thus 44.4 % (24/53). The only embryo failed to provide a confirmed diagnosis showed poor qPCR signal, possibly due to a low amount of or degraded DNA in the biopsied sample. FISH diagnosis for that embryo was negative for numerical disorders involving chromosome 13, 18, 21, X, and Y. The remaining cells of this biopsied embryo were sent for array CGH and the diagnosis was trisomy 22. Clinical verification A total of 72 embryos at Day 5/6 blastocyst stage from 13 patients were biopsied and assessed, and all (100 %) of them were successfully diagnosed (Table 1 and Additional file 1: Table S1). Twenty-eight (38.9 %) of these embryos were aneuploid, which included 4 monosomy 13, 5 monosomy 21, 7 trisomy 13, 1 trisomy 18, 9 trisomy 21, 1 monosomy 13 + trisomy 18, and 1 monosomy 18 + monosomy 21. The remaining 44 (61.1 %) embryos were euploidy, of which 28 were transferred into the 13 patients in 13 cycles. The average number of the embryos being transferred is 2.15, actually only 1–2 embryos were transferred in all women of the Group B (non-AMA group, n = 10). The only 2 women who were transferred for 4 embryos are both in the Group A (AMA group, n = 3). The transfer procedure follows the medical rule of Taiwan that the upper limit of the number of the embryos being transferred is 4. These transfers achieved 8 pregnancies, and therefore the clinical pregnancy rate was 61.5 % (8/13) per cycle. However, one early abortion was noted and the ongoing pregnancy rate up to the second trimester was 53.8 % (7/13) per transfer cycle. There were no monozygotic twinning and a total of 11 sacs were noted. Three women who were transferred for 2 embryos in the Group B had singleton pregnancies, and two women who were transferred for 2 embryos in the Group B and 2 women who were transferred for 4 embryos in the Group A had dizygotic twin pregnancies (Additional file 1: Table S1). The sustained implantation rate was thus 39.3 % (11/28) per transferred fresh embryo (Table 1). Totally, there are 2 of 3 women in Group A had ongoing pregnancies. Six women in Group B had clinical pregnancies, and 5 of them had sustained ongoing pregnancies. There are no statistical significant differences noted between these two groups regarding aneuploidy (Chi square test, p = 0.094), clinical pregnancy (Fisher's exact test, p = 1), and ongoing pregnancy (Fisher's exact test, p = 1). Array CGH for the only abortion case revealed the karyotype was 46,XY. Table 1. Clinical outcomes of PGS by qPCR with fresh embryo transfer (FET) verification series Discussion The major difficulty of comprehensive chromosome screening (CCS) by qPCR for PGS is to devise a primer set to successfully amplify the molecular markers being selected in a single experiment by optimizing the conditions for melting temperatures and experimental time intervals. In this study we successfully adopted a smart design called LNA methodology [20], [22]–[24] for qPCR PGS and verified the strategy by achieving a favorable ongoing pregnancy rate of 53.8 %. We first validated the genotyping platform we devised by achieving an almost 100 % successful diagnosis rate (98.1 %), and then verified the whole protocol of qPCR PGS followed by FET. Since the sample size is small (n = 13 cycles), it is not surprising that we failed to observe significant difference regarding the aneuploidy rates between the AMA (n = 3) and the non-AMA group (n = 10) in our patients. Meanwhile, the aneuploidy rate in our series (38.9 %) is apparently much lower than the rates observed by 24-chromosome qPCR or array CGH (more than 60 %), which is obviously due to the fact that we only selected a limited set of common aneuploidies that can ensue live births in humans [14], [15]. It has been a routine practice in our settings that PGS by FISH at Day 3 cleavage-stage embryos since 2005 and it is ethical for us to offer our patients who opted for this alternative protocol a strategy which has an advantage of an established less detrimental effect upon implantation potential by changing the timing of biopsy from Day 3 to Day 5/6 and the same repertoire of common aneuploidies being tested (but simply using qPCR instead of FISH) to enable FET[9]. The reason only trisomy 13, 18, 21, and numerical disorders involving X and Y were tested in our settings is because these are the only aneuploidies that may result in live births in humans. For people who opted for this strategy and came to our clinics, their major concern is more focused at aneuploidies instead of live birth rates. In addition, it is well recognized that despite fewer embryos would be available for transfer if being cultured in vitro onto the blastocyst stage when compared with the cleavage-stage embryos, the disadvantage may be offset by the fact that the aneuploidy rate of the blastocysts is much lower than of the cleavage-stage embryos. An observational study had reported that karyotypic evolution did exist in early human embryogenesis, in which some cleavage stage embryos classified as aneuploidy by FISH eventually developed into euploid blastocysts if by SNP array[25]. A recent randomized study even pointed out that FISH-based PGS conferred advantages at women who are in the AMA group, but not in those with repeated implantation failures group, as long as the biopsy was performed at the blastocyst stage [5]. A recent randomized trial (the BEST trial) had demonstrated that qPCR PGS CCS followed by single embryo transfer (SET) can enhance the feasibility of SET by selecting a single euploid embryo with high reproductive potential without compromising the delivery rates when compared with transfer of two unselected embryos, which instead resulted in more multiple pregnancies and the associated complications such as preterm delivery, low birth weight, and NICU (Neonatal Intensive Care Unit) admission [26]. We admit our pilot verification study had suffered from the fact that only common aneuploidies of five chromosome pairs (13, 18, 21, X, and Y) were screened and apparently it is the reason that the implantation rate per embryo in our study (39.3 %) is much lower than that in the BEST trial (69 %) [26]. However, our patients were all informed consented and knew the disadvantage before joining the study, and the limitations of the local setting were clearly disclosed to the patients. The patients were free to choose another PGS protocol by array CGH (Option 2, please refer to Patients and methods section) instead. Conclusions The merit of our work is that this is the first effort from research groups other than the original group who proposed the qPCR CCS (Reproductive Medicine Associates of New Jersey, Morristown, NJ, USA) trying to validate and verify this novel genotyping platform which was developed in-house by ourselves. Despite our results did not have matched controls such as those cycles receiving IVF but without PGS, the ongoing pregnancy rate is similar to a previous PGD series from our setting (53.8 % versus 50 %, please refer to Chang et al., 2013 [27]), indicating this protocol is clinically feasible. Future efforts will be made upon expanding the repertoire of the PGS by qPCR CCS to all 24 chromosomes, as well as carefully-designed randomized controlled trials to compare this genotyping platform with others (including no-screening, array CGH, and NGS). Patients and methods Design and selection of molecular markers for qPCR PGS An in-house screening system by dual-color qPCR was developed with specific primers for the targeted loci (situated at chromosome 13, 18, 21, X, and Y) and one internal control locus (situated at chromosome 1, the reason we chose chromosome 1 as control is because it is the least found trisomy in humans, see Wang et al., 2014 [21]). A total of 16 targeted loci and one control locus were selected. For the detailed information about the targeted loci, please refer to Table 2. Specific primers for amplification of each locus were designed by using the software Oligo 6.71 (Molecular Biology Insights, Colorado, USA), and all primer sets are flanked on each side of the exon-intron boundary to avoid possible mRNA interference. The applications of LNA-modified probes are well-established for quantifying levels of gene expression with an advantage of reducing complexity [22], [24], but has not been applied to the detection of chromosomal copy number alterations. Two kinds of LNA-modified probes, labeled with FAM™ and HEX™ (Integrated DNA Technologies, Iowa, USA) at the 5’-end, were used for quantifying the genomic copy numbers of targeted loci and control. Table 2. Summary of the targeted regions for screening of common aneuploidies Validation of qPCR in detecting common aneuploidies Each 5 cells separated from the cell lines of known common aneuploidies (including trisomy 21, trisomy 13, trisomy 18, 47,XXY, and 45,X) were processed by cell lysis of proteinase K, and the products were subjected to a 50-μL reaction volume of multiplex nested-PCR amplication for 18 cycles using an Applied Biosystems Veriti thermal cycler (Life Technologies, California, USA), and then the PCR products were purified using Agencourt AMPure XP system (Beckman Coulter, California, USA). Dual color hydrolysis probe assays were performed in triplicate to normalize and simpify calculation and to evaluate chromosomal copies, using Lightcycler 480 probes Master (Roche, Mannheim, Germany), a 20-μL reaction volume, a 96-well plate, and a 7 Light Cycler 480 Real-Time PCR System, as recommended by the supplier (Roche, Mannheim, Germany). Each well contains a particular target, and a common control reaction. A unique method of the standard delta delta threshold cycle (ΔΔCp) method was used for relative quantification. In our experiments, the Cp variation of all HEX™ reactions obtained for each well of the same sample will be controlled and ranged in less than 0.2, indicating the test sample was evenly distributed to each well. Each chromosome-specific ΔCp was calculated from the Cp of the FAM™ reactions targeting a specific chromosome minus the control Cp of the HEX™ reactions targeting the chromosome 1 within the same well. The same process was applied to individually determine the ΔCp for each targeted chromosome of the test sample, including reference set of normal male cell lines [BCRC number: 08C0011, 08C0012, 08C0013, 08C0021 and 08C0025]. Each chromosome- specific ΔCp was then normalized to the average chromosome-specific ΔCp values derived from the same evaluation of the reference set, which had been confirmed by FISH method. The calibrated chromosome-specific ΔCp values were used to calculate fold change by considering the ΔΔCp values as the negative exponent of 2, as previously described [18], [23]. The methodology was designed to specifically identify whole-chromosome but not segmental aneuploidy. The flowchart and diagram of the in-house qPCR PGS system were illustrated in Fig. 1. This qPCR was capable of accurate aneuploidy screening in 4 h, which allowed rapid evaluation of the trophectoderm biopsies and therefore provided a feasible opportunity for subsequent FET. thumbnailFig. 1. The diagram of the in-house qPCR PGS system. The flowchart of (a) detection of common aneuploidies, and (b) signal normalization and data analysis Pre-clinical validation upon surplus frozen embryos Fifty-four thawed frozen embryos at blastocyst stage were biopsied and sent for qPCR and FISH analyses. These embryos were retrieved from the surplus frozen embryos of couples who already conceived and would be discarded if not being investigated for research purpose. In those embryos diagnosed with common chromosome aneuploidies, array CGH was used to confirm the diagnoses. Clinical verification for fresh embryos In our setting we used to offer two PGS protocols. One option (Option 1) is Day 3 biopsy followed by PGS with FISH and FET; the other option (Option 2) is Day 5/6 trophectoderm biopsy followed by PGS with array CGH and frozen embryo transfer (because array CGH takes time and FET was not feasible at the study period). Only patients who had history of failed IVF (without PGS) for at least once and who opted for Option 1 were given the chance of joining this study as an alternative. All patients chose to join this study were informed consented and their autonomy was fully respected. They could choose to withdraw from the study at any time during the study period and were fully aware of the alternatives, including sticking to Option 1 or instead chose Option 2 without joining the study. During July to October of 2014, 13 infertile couples were enrolled. Among these 13 patients, 3 of them had AMA (37, 43, 45 years old respectively). The mean age of the total 13 patients was 34.1 years. No confounding factors that may affect implantation such as immune aberrations (antiphospholipid antibody syndrome in the mother), balanced translocation carriers (in both couples), and thrombophilias (protein C/S/antithrombin III deficiency) existed in these couples when they were enrolled. Clinical pregnancy was defined as positive urine HCG. Ongoing pregnancy rate (per cycle) was defined as those pregnancies proceeding into second trimester (and for each embryo being transferred, sustained implantation rate was used). 13 couples were classified as those whose age is older or equal to 35 years (Group A), and those whose age is less than 35 years (Group B). Chi square test or Fisher’s exact test was used to compare the reproductive outcomes between the groups regarding the rate of aneuploidy, clinical pregnancy, and ongoing pregnancy. Notably according to the regulations of Taiwan government, the patients would not know the results of the fetal sex. Even in aneuploidies involving sex chromosomes (specifically 47,XXY and 45,X), the results would not be disclosed to patients and only “aneuploidy” would be told to the patients and the aneuploid embryos would not be selected for transfer. In those embryos diagnosed to have chromosomal aneuploidies by qPCR, array CGH was used to confirm the diagnoses. Abbreviations AMA: Advanced maternal age CCS: Comprehensive chromosome screening CGH: Comparative genomic hybridization FET: Fresh embryo transfer FISH: Fluorescence in situ hybridization IVF: in vitro fertilization LNA: Locked nucleic acid NGS: Next generation sequencing PGS: Preimplantation genetic screening qPCR: Quantitative real-time polymerase chain reaction SET: Single embryo transfer SNP: Single nucleotide polymorphism Competing interests The authors declare that they have no competing interests. Authors’ contributions YSY, HFC, SPC, FPT, MC designed the study. SPC, GCM, MC developed the novel in-house genotyping platform. SPC, GCM, WHL, CFL, MC did the experiments. YSY, HFC, FPT, CHW, HDT, MC recruited the patients. YSY, HFC, GCM, THL, MC analyzed the results. YSY, SPC, GCM, WHL, MC wrote the paper. All authors read and approved the final manuscript. Additional file Additional file 1: Table S1.. Summary of the pre-clinical validation (for 54 surplus frozen embryos) and clinical verification (for 13 patients with 54 embryos) of PGS by qPCR. Format: DOCX Size: 32KB Download fileOpen Data Acknowledgements This study is partly funded by a grant from Changhua Christian Hospital, Taiwan to Ming Chen (103-CCH-IST-006). The authors are grateful for the genuine support and advice from Dr. Shou-Jen Kuo, the Superintendent of Changhua Christian Hospital, who also participated in the development of the novel in-house qPCR CCS system. References Treff NR, Scott RT. Methods for comprehensive chromosome screening of oocytes and embryos: capabilities, limitations, and evidence of validity. 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• 2014: 胚胎切片全基因放大檢測會增加試管嬰兒成功率嗎？一個案例報告ccs T小姐因為年紀因素，因此到博元婦產科進行試管嬰兒，她了解在懷孕之後需要做羊水穿刺，因此要求在囊胚期胚胎階段進行胚胎全基因放大檢測，我們執行胚胎切片送全基因放大檢測，等2個禮拜有了報告，有一個胚胎染色體異常（見下面照片）過程為：打排卵針、取卵手術、第五天胚胎切片囊胚期胚胎階段進行胚胎全基因放大檢測、冷凍胚胎，等2個禮拜報告出來之後，再植入染色體正常的胚胎，目前已經成功懷孕2胞胎，你現在看到的雙胞胎的影片就是她的寶寶，恭喜她。 試管嬰兒進入第三代，我們進行胚胎切片著床前診斷，分成兩大部分： 1.PGD找出健康寶寶不帶疾病，這是有特地單一基因疾病要排除，簡單地說，父親或母親有遺傳性疾病，不想遺傳給下一代子女，想要生健康寶寶，這叫做PGD胚胎著床前基因診斷。 2.胚胎全基因放大檢測，等於找出染色體全部正常的胚胎才植入子宮，目前證據顯示可以增加試管嬰兒成功率、胚胎著床率、試管嬰兒懷孕率及活產率，目前胚胎切片囊胚期胚胎階段進行胚胎全基因放大檢測用微陣列全基因放大（array-CGH）的報告要兩個禮拜，希望將來有4小時快速基因檢測（qPCR），就可以在第5天切片，第5天植入或第6天植入胚胎，根據美國紐澤西洲理查醫師研究團隊，這可以增加試管嬰兒成功率，2個染色體正常的胚胎植入，活產率是84.7%。 https://www.youtube.com/watch?v=-43SL4w8MKw <iframe width="560" height="315" src="//www.youtube.com/embed/-43SL4w8MKw" frameborder="0" allowfullscreen></iframe> 胚胎切片全基因放大檢測會增加試管嬰兒成功率嗎？一個案例報告ccs T小姐因為年紀因素，因此到博元婦產科進行試管嬰兒，她了解在懷孕之後需要做羊水穿刺，因此要求在囊胚期胚胎階段進行胚胎全基因放大檢測，我們執行胚胎切片送全基因放大檢測，等2個禮拜有了報告，有一個胚胎染色體異常（見下面照片）過程為：打排卵針、取卵手術、第五天胚胎切片囊胚期胚胎階段進行胚胎全基因放大檢測、冷凍胚胎，等2個禮拜報告出來之後，再植入染色體正常的胚胎，目前已經成功懷孕2胞胎，你現在看到的雙胞胎的影片就是她的寶寶，恭喜她。 試管嬰兒進入第三代，我們進行胚胎切片著床前診斷，分成兩大部分： 1.PGD找出健康寶寶不帶疾病，這是有特地單一基因疾病要排除，簡單地說，父親或母親有遺傳性疾病，不想遺傳給下一代子女，想要生健康寶寶，這叫做PGD胚胎著床前基因診斷。 2.胚胎全基因放大檢測，等於找出染色體全部正常的胚胎才植入子宮，目前證據顯示可以增加試管嬰兒成功率、胚胎著床率、試管嬰兒懷孕率及活產率，目前胚胎切片囊胚期胚胎階段進行胚胎全基因放大檢測用微陣列全基因放大（array-CGH）的報告要兩個禮拜，希望將來有4小時快速基因檢測（qPCR），就可以在第5天切片，第5天植入或第6天植入胚胎，根據美國紐澤西洲理查醫師研究團隊，這可以增加試管嬰兒成功率，2個染色體正常的胚胎植入，活產率是84.7%。 https://www.youtube.com/watch?v=-43SL4w8MKw ( 知識學習｜健康 )
• 2014: 無精症：XXY症候群，經博元婦產科精子銀行,4小時快篩胚胎染色體試管嬰兒,一次成功懷孕 qPCR bank 先生無精症，47XXY症候群，經博元婦產科精子銀行試管嬰兒一次成功懷孕qPCR， Y小姐跟先生結婚兩年，一直都無法受孕， 到博元婦產科檢查才知道，先生是屬於男性無造精功能的無精症， 經過抽血荷爾蒙染色體的層層分析證實， 他無法用打針的方式造出精子， 因此進行精子銀行作業，一次精子銀行試管嬰兒就成功， 因為他們的基因染色體異常，所以他們在做試管嬰兒的胚胎時， 在第5天囊胚期，進行胚胎切片， 植入胚胎前一天做胚胎切片，進行4小時快篩胚胎染色體， 4小時就有報告， 我們第六天植入胚胎，現在已經證實成功懷孕，並且懷孕5週，她在驗孕日當天，他的懷孕抽血值：人類絨毛膜刺激激素指數是367.6， 雖然目前看到只有一個胚囊，可以預期在6週也許可以看到2個胚胎，恭喜！ 使用 (1)精子銀行 (2)胚胎4小時快篩染色體，經過檢查，染色體正常，植入胚胎成功懷孕
• 2014: 不孕症六年，在彰化博元婦產科只做一次試管嬰兒，冷凍胚胎成功懷孕 fet 不孕症六年，在彰化博元婦產科做試管嬰兒，冷凍胚胎成功懷孕， C小姐她跟先生結婚六年，都一直不孕，到處求醫也都無法一圓子望， 最近至博元婦產科，一開口就說要求做試管嬰兒， 她的理由就是在別家醫院已經做過很多次人工授精， 但都不能一圓子望， 我們進行試管嬰兒取卵當天， 發現她的動情指數偏高， 在擔心卵巢過度刺激的風險的情況下， 我們進行 冷凍胚胎， 下一個月 植入胚胎， 她植入胚胎的囊胚， 是非常的優， 我們植入胚胎的數量符合她的年齡， 她驗孕當天的β-HCG是322，預期大概有1胞胎至2胞胎， 恭喜她！在不孕症六年之後， 在彰化博元婦產科， 以只有一次的試管嬰兒的冷凍胚胎， 解凍胚胎 進行試管嬰兒成功懷孕。今天來做超音波好像五週三胞胎見超音波照片；
• 2014: 不孕症六年，在彰化博元婦產科只做一次試管嬰兒，冷凍胚胎成功懷孕 fet 不孕症六年，在彰化博元婦產科做試管嬰兒，冷凍胚胎成功懷孕， C小姐她跟先生結婚六年，都一直不孕，到處求醫也都無法一圓子望， 最近至博元婦產科，一開口就說要求做試管嬰兒， 她的理由就是在別家醫院已經做過很多次人工授精， 但都不能一圓子望， 我們進行試管嬰兒取卵當天， 發現她的動情指數偏高， 在擔心卵巢過度刺激的風險的情況下， 我們進行 冷凍胚胎， 下一個月 植入胚胎， 她植入胚胎的囊胚， 是非常的優， 我們植入胚胎的數量符合她的年齡， 她驗孕當天的β-HCG是322，預期大概有1胞胎至2胞胎， 恭喜她！在不孕症六年之後， 在彰化博元婦產科， 以只有一次的試管嬰兒的冷凍胚胎， 解凍胚胎 進行試管嬰兒成功懷孕。今天來做超音波好像五週三胞胎見超音波照片；
• 2014: 無精症：XXY症候群，經博元婦產科精子銀行,4小時快篩胚胎染色體試管嬰兒,一次成功懷孕 qPCR bank 先生無精症，47XXY症候群，經博元婦產科精子銀行試管嬰兒一次成功懷孕qPCR， Y小姐跟先生結婚兩年，一直都無法受孕， 到博元婦產科檢查才知道，先生是屬於男性無造精功能的無精症， 經過抽血荷爾蒙染色體的層層分析證實， 他無法用打針的方式造出精子， 因此進行精子銀行作業，一次精子銀行試管嬰兒就成功， 因為他們的基因染色體異常，所以他們在做試管嬰兒的胚胎時， 在第5天囊胚期，進行胚胎切片， 植入胚胎前一天做胚胎切片，進行4小時快篩胚胎染色體， 4小時就有報告， 我們第六天植入胚胎，現在已經證實成功懷孕，並且懷孕5週，她在驗孕日當天，他的懷孕抽血值：人類絨毛膜刺激激素指數是367.6， 雖然目前看到只有一個胚囊，可以預期在6週也許可以看到2個胚胎，恭喜！ 使用 (1)精子銀行 (2)胚胎4小時快篩染色體，經過檢查，染色體正常，植入胚胎成功懷孕
• 2014: 陰道痙攣無性愛　卻懷了雙胞胎--- 壹週刊 nextmag: -----彰化博元婦產科蔡鋒博將在該診所因陰道痙攣進行人工生殖經驗的43例患者進行分析，發現有19個懷孕成功，其中24位人工授精病人中有8位成功懷孕，19位試管嬰兒病人中有11位懷孕成功。 ----- 彰化博元婦產科最近幫一對結婚10年無法做愛的夫妻完成人工授孕。院長蔡鋒博說，夫妻初診當天就進入試管嬰兒療程，做什麼都要麻醉，包括陰道超音波、取卵手術、植入胚胎手術、子宮鏡，還好一次就成功懷了雙胞胎，目前已12週 ------彰化博元婦產科最近幫一對從台北南下求子的陰道痙攣夫婦，9年來通通沒有性經驗，可以說是童貞夫人，麻醉後要做人工授精才發現，病人的處女膜是完整的，還好人工授精的管子非常細，就從處女膜的洞口插進去，也讓患者成功懷孕了，生完產後，處女膜還是完整。 。內置圖片 1 http://www.nextmag.com.tw/breaking-news/life/20140922/8385726 陰道痙攣無性愛卻懷了雙胞胎｜Next Magazine - 壹週刊 www.nextmag.com.tw/breaking-news/life/20140922/8385726 2 陰道痙攣是一個非常特殊又讓人難以啟齒的疾病，發生機率不到2%，這類患者無法性行為，若結了婚想生孩子怎麼辦，就得利用人工生殖來完成。 2014年09月22日 16:18 http://www.nextmag.com.tw/breaking-news/life/20140922/8385726 http://www.nextmag.com.tw/breaking-news/life/20140922/8385726 Facebook Plurk Google Plus 嚴重的陰道痙攣無法性行為，想生孩子就得靠人工授精來完成傳宗接代的任務。（資料照片） 嚴重的陰道痙攣無法性行為，想生孩子就得靠人工授精來完成傳宗接代的任務。（資料照片） 陰道痙攣是一個非常特殊又讓人難以啟齒的疾病，發生機率不到2%，這類患者無法性行為，若結了婚想生孩子怎麼辦，就得利用人工生殖來完成。彰化博元婦產科最近幫一對結婚10年無法做愛的夫妻完成人工授孕。院長蔡鋒博說，夫妻初診當天就進入試管嬰兒療程，做什麼都要麻醉，包括陰道超音波、取卵手術、植入胚胎手術、子宮鏡，還好一次就成功懷了雙胞胎，目前已12週。 日前也有一名從台北南下求子的陰道痙攣夫婦，9年來通通沒有性經驗，可以說是童貞夫人，麻醉後要做人工授精才發現，病人的處女膜是完整的，還好人工授精的管子非常細，就從處女膜的洞口插進去，也讓患者成功懷孕了，生完產後，處女膜還是完整。 蔡鋒博將在該診所因陰道痙攣進行人工生殖經驗的43例患者進行分析，發現有19個懷孕成功，其中24位人工授精病人中有8位成功懷孕，19位試管嬰兒病人中有11位懷孕成功。 蔡鋒博說，陰道痙攣在治療上，可以先進行精神科的專業諮詢；再來可由物治療來幫忙，如由小到大的人工陰莖或鴨嘴，或藉由藥物治療，如中樞神經系統肌肉鬆弛劑；若都沒效，還是無法正常行房，那麼人工生殖就是最後手段。（撰文：蔡怡真）女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四， 中視新聞》女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 國內超過萬名女性患陰道痙攣症，導致無法順利行房甚至受孕。一名三十二歲女子就因為這­樣，婚後十年從未行房、仍然是處女。因此醫師只能嘗試試管嬰兒療程，讓她順利的懷了一­對雙胞胎。 http://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="960" height="720" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> http://www.youtube.com/watch?v=j9u1lgHwXjU <iframe width="960" height="720" src="//www.youtube.com/embed/j9u1lgHwXjU" frameborder="0" allowfullscreen></iframe> 中視新聞》女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 【中視新聞】 女陰道痙攣無法受孕 博元婦產科嘗試試管一舉懷雙胞 20140922 https://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="420" height="315" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="480" height="360" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=blhiir8PldY <iframe width="1280" height="720" src="//www.youtube.com/embed/blhiir8PldY" frameborder="0" allowfullscreen></iframe> 收治病例的博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 -- 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四， 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師
• 2014: 陰道痙攣無性愛　卻懷了雙胞胎--- 壹週刊 nextmag: -----彰化博元婦產科蔡鋒博將在該診所因陰道痙攣進行人工生殖經驗的43例患者進行分析，發現有19個懷孕成功，其中24位人工授精病人中有8位成功懷孕，19位試管嬰兒病人中有11位懷孕成功。 ----- 彰化博元婦產科最近幫一對結婚10年無法做愛的夫妻完成人工授孕。院長蔡鋒博說，夫妻初診當天就進入試管嬰兒療程，做什麼都要麻醉，包括陰道超音波、取卵手術、植入胚胎手術、子宮鏡，還好一次就成功懷了雙胞胎，目前已12週 ------彰化博元婦產科最近幫一對從台北南下求子的陰道痙攣夫婦，9年來通通沒有性經驗，可以說是童貞夫人，麻醉後要做人工授精才發現，病人的處女膜是完整的，還好人工授精的管子非常細，就從處女膜的洞口插進去，也讓患者成功懷孕了，生完產後，處女膜還是完整。 。內置圖片 1 http://www.nextmag.com.tw/breaking-news/life/20140922/8385726 陰道痙攣無性愛卻懷了雙胞胎｜Next Magazine - 壹週刊 www.nextmag.com.tw/breaking-news/life/20140922/8385726 2 陰道痙攣是一個非常特殊又讓人難以啟齒的疾病，發生機率不到2%，這類患者無法性行為，若結了婚想生孩子怎麼辦，就得利用人工生殖來完成。 2014年09月22日 16:18 http://www.nextmag.com.tw/breaking-news/life/20140922/8385726 http://www.nextmag.com.tw/breaking-news/life/20140922/8385726 Facebook Plurk Google Plus 嚴重的陰道痙攣無法性行為，想生孩子就得靠人工授精來完成傳宗接代的任務。（資料照片） 嚴重的陰道痙攣無法性行為，想生孩子就得靠人工授精來完成傳宗接代的任務。（資料照片） 陰道痙攣是一個非常特殊又讓人難以啟齒的疾病，發生機率不到2%，這類患者無法性行為，若結了婚想生孩子怎麼辦，就得利用人工生殖來完成。彰化博元婦產科最近幫一對結婚10年無法做愛的夫妻完成人工授孕。院長蔡鋒博說，夫妻初診當天就進入試管嬰兒療程，做什麼都要麻醉，包括陰道超音波、取卵手術、植入胚胎手術、子宮鏡，還好一次就成功懷了雙胞胎，目前已12週。 日前也有一名從台北南下求子的陰道痙攣夫婦，9年來通通沒有性經驗，可以說是童貞夫人，麻醉後要做人工授精才發現，病人的處女膜是完整的，還好人工授精的管子非常細，就從處女膜的洞口插進去，也讓患者成功懷孕了，生完產後，處女膜還是完整。 蔡鋒博將在該診所因陰道痙攣進行人工生殖經驗的43例患者進行分析，發現有19個懷孕成功，其中24位人工授精病人中有8位成功懷孕，19位試管嬰兒病人中有11位懷孕成功。 蔡鋒博說，陰道痙攣在治療上，可以先進行精神科的專業諮詢；再來可由物治療來幫忙，如由小到大的人工陰莖或鴨嘴，或藉由藥物治療，如中樞神經系統肌肉鬆弛劑；若都沒效，還是無法正常行房，那麼人工生殖就是最後手段。（撰文：蔡怡真）女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四， 中視新聞》女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 國內超過萬名女性患陰道痙攣症，導致無法順利行房甚至受孕。一名三十二歲女子就因為這­樣，婚後十年從未行房、仍然是處女。因此醫師只能嘗試試管嬰兒療程，讓她順利的懷了一­對雙胞胎。 http://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="960" height="720" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> http://www.youtube.com/watch?v=j9u1lgHwXjU <iframe width="960" height="720" src="//www.youtube.com/embed/j9u1lgHwXjU" frameborder="0" allowfullscreen></iframe> 中視新聞》女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 【中視新聞】 女陰道痙攣無法受孕 博元婦產科嘗試試管一舉懷雙胞 20140922 https://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="420" height="315" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="480" height="360" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=blhiir8PldY <iframe width="1280" height="720" src="//www.youtube.com/embed/blhiir8PldY" frameborder="0" allowfullscreen></iframe> 收治病例的博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 -- 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四， 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師
• 2014: 太神奇！婚後產子仍是「童貞夫人」 華人健康網／記者黃曼瑩／台北報導-2014年09月22日 上午11:30 http://history.n.yam.com/top1health/healthy/20140922/20140922569504.html 中視新聞》女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 http://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="960" height="720" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> http://www.youtube.com/watch?v=j9u1lgHwXjU <iframe width="960" height="720" src="//www.youtube.com/embed/j9u1lgHwXjU" frameborder="0" allowfullscreen></iframe> cteditor cteditor 內置圖片 1 字級： 小 中 大 特 列印 轉寄 分享 太神奇！婚後產子仍是「童貞夫人」 arrow-left 1/2 arrow_right 點選放大 女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 【中視新聞】 女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 20140922 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 http://www.chinatimes.com/realtimenews/20140923001538-260405 http://www.youtube.com/watch?v=j9u1lgHwXjU <iframe width="420" height="315" src="//www.youtube.com/embed/j9u1lgHwXjU" frameborder="0" allowfullscreen></iframe> 2014年09月23日 07:44 金維中、吳烈安、曾新中 小字型 中字型 大字型 185點閱 2/10我要評比 0 影音新聞來源/中視提供 國內有超過萬名女性罹患陰道痙攣症，島至無法行房順利懷孕，嚴重一點的甚至檢查都得麻醉才能進行，更有大半數婦女因為害羞，一直不願意就診。 一名女子就是罹患重度痙攣症，結婚十幾年來從沒和丈夫行房過，但因為想要子女，醫師只能採用試管嬰兒，由於他的病情嚴重，從超音波檢查到取卵，一切都要麻醉後，才能進行。 最後女子終於順利懷的一對雙胞胎，醫師也表示，這種疾病在婦女監並不罕見，因此如果有相關病證，還是及早治療，才不會延誤到夫妻間的幸福。【中視新聞】 女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 20140922 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="480" height="360" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=blhiir8PldY <iframe width="1280" height="720" src="//www.youtube.com/embed/blhiir8PldY" frameborder="0" allowfullscreen></iframe> 收治病例的博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 -- 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四， 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 你相信嗎？1名32歲罹患嚴重陰道痙攣症的女子結婚10年，竟然丈夫從來都沒有「碰」過太太任何一次，更神奇的是，女子現已懷孕12周，將來剖腹產後仍是完璧之身，成為名副其實的「童貞夫人」！一項由國內婦產科醫師發表於台灣生殖醫學會年會上的特殊案例，令人嘖嘖稱奇。 台灣罹患陰道痙攣症(vaginismus)的婦女比例大約有1％，陰道痙攣症會嚴重影響女人進行性生活的活動無法正常進行。 婦產科醫師蔡鋒博表示，台灣罹患陰道痙攣症(vaginismus)的婦女比例大約有1％，陰道痙攣症會嚴重影響女人進行性生活的活動無法正常進行，如陰道插入：包括正常的性交，或插入衛生棉條，或使用月經杯，甚至一般婦產科醫師的內診，或使用鴨嘴做陰道檢查，都無法順利進行。 嚴重陰道痙攣症　易導致不孕 醫學上研究發現，證實患有嚴重陰道痙攣症病人，每當有外物靠近外陰部，或僅憑自己幻想可能有這樣子的動作預期，就會引起陰道肌肉群不自主強烈收縮，引發這種陰道肌肉強烈收縮的因素，包括：性交，會讓性交變的很疼痛，或根本無法正常進行，也因此使得男女無法正常性交，性伴侶無法正常在陰道射精，自然就衍生後續不孕症的問題。 這1名求診的婦女嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，所以產後仍是處女，乖可以說是難得一見的「童貞夫人」。 如果有陰道痙攣症要求子，儘早做治療要趁年輕。 陰道痙攣原因多　心理問題為主 引起陰道痙攣的原因主要與心理因素有關，多數患者由於對性知識的缺乏，對性交的極端恐懼與焦慮而產生，遭受過性暴力或外陰和陰道有創傷史的女性也易產生。以及夫妻感情不合，對配偶的不滿都有影響。 【醫師小叮嚀】： 蔡鋒博醫師表示，如果有陰道痙攣症要求子，儘早做治療要趁年輕，如果經由以下方式都失敗，包括： 1.精神科專業技巧的幫助諮詢。 2.物理治療，如由小到大的人工陰莖或鴨嘴。 3.藥物治療，如中樞神經系統肌肉鬆弛劑。 以上三個方法都失敗的話，人工助孕包括人工授精跟試管嬰兒，是重度陰道痙攣症求子的最後一張王牌！ 文章連結 http://www.top1health.com/Article/12/20293
• 2014: 太神奇！婚後產子仍是「童貞夫人」 華人健康網／記者黃曼瑩／台北報導-2014年09月22日 上午11:30 http://history.n.yam.com/top1health/healthy/20140922/20140922569504.html 中視新聞》女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 http://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="960" height="720" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> http://www.youtube.com/watch?v=j9u1lgHwXjU <iframe width="960" height="720" src="//www.youtube.com/embed/j9u1lgHwXjU" frameborder="0" allowfullscreen></iframe> cteditor cteditor 內置圖片 1 字級： 小 中 大 特 列印 轉寄 分享 太神奇！婚後產子仍是「童貞夫人」 arrow-left 1/2 arrow_right 點選放大 女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 【中視新聞】 女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 20140922 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 http://www.chinatimes.com/realtimenews/20140923001538-260405 http://www.youtube.com/watch?v=j9u1lgHwXjU <iframe width="420" height="315" src="//www.youtube.com/embed/j9u1lgHwXjU" frameborder="0" allowfullscreen></iframe> 2014年09月23日 07:44 金維中、吳烈安、曾新中 小字型 中字型 大字型 185點閱 2/10我要評比 0 影音新聞來源/中視提供 國內有超過萬名女性罹患陰道痙攣症，島至無法行房順利懷孕，嚴重一點的甚至檢查都得麻醉才能進行，更有大半數婦女因為害羞，一直不願意就診。 一名女子就是罹患重度痙攣症，結婚十幾年來從沒和丈夫行房過，但因為想要子女，醫師只能採用試管嬰兒，由於他的病情嚴重，從超音波檢查到取卵，一切都要麻醉後，才能進行。 最後女子終於順利懷的一對雙胞胎，醫師也表示，這種疾病在婦女監並不罕見，因此如果有相關病證，還是及早治療，才不會延誤到夫妻間的幸福。【中視新聞】 女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 20140922 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="480" height="360" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=blhiir8PldY <iframe width="1280" height="720" src="//www.youtube.com/embed/blhiir8PldY" frameborder="0" allowfullscreen></iframe> 收治病例的博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 -- 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四， 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 你相信嗎？1名32歲罹患嚴重陰道痙攣症的女子結婚10年，竟然丈夫從來都沒有「碰」過太太任何一次，更神奇的是，女子現已懷孕12周，將來剖腹產後仍是完璧之身，成為名副其實的「童貞夫人」！一項由國內婦產科醫師發表於台灣生殖醫學會年會上的特殊案例，令人嘖嘖稱奇。 台灣罹患陰道痙攣症(vaginismus)的婦女比例大約有1％，陰道痙攣症會嚴重影響女人進行性生活的活動無法正常進行。 婦產科醫師蔡鋒博表示，台灣罹患陰道痙攣症(vaginismus)的婦女比例大約有1％，陰道痙攣症會嚴重影響女人進行性生活的活動無法正常進行，如陰道插入：包括正常的性交，或插入衛生棉條，或使用月經杯，甚至一般婦產科醫師的內診，或使用鴨嘴做陰道檢查，都無法順利進行。 嚴重陰道痙攣症　易導致不孕 醫學上研究發現，證實患有嚴重陰道痙攣症病人，每當有外物靠近外陰部，或僅憑自己幻想可能有這樣子的動作預期，就會引起陰道肌肉群不自主強烈收縮，引發這種陰道肌肉強烈收縮的因素，包括：性交，會讓性交變的很疼痛，或根本無法正常進行，也因此使得男女無法正常性交，性伴侶無法正常在陰道射精，自然就衍生後續不孕症的問題。 這1名求診的婦女嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，所以產後仍是處女，乖可以說是難得一見的「童貞夫人」。 如果有陰道痙攣症要求子，儘早做治療要趁年輕。 陰道痙攣原因多　心理問題為主 引起陰道痙攣的原因主要與心理因素有關，多數患者由於對性知識的缺乏，對性交的極端恐懼與焦慮而產生，遭受過性暴力或外陰和陰道有創傷史的女性也易產生。以及夫妻感情不合，對配偶的不滿都有影響。 【醫師小叮嚀】： 蔡鋒博醫師表示，如果有陰道痙攣症要求子，儘早做治療要趁年輕，如果經由以下方式都失敗，包括： 1.精神科專業技巧的幫助諮詢。 2.物理治療，如由小到大的人工陰莖或鴨嘴。 3.藥物治療，如中樞神經系統肌肉鬆弛劑。 以上三個方法都失敗的話，人工助孕包括人工授精跟試管嬰兒，是重度陰道痙攣症求子的最後一張王牌！ 文章連結 http://www.top1health.com/Article/12/20293
• 2014: 太神奇！婚後產子仍是「童貞夫人」 華人健康網／記者黃曼瑩／台北報導-2014年09月22日 上午11:30 http://history.n.yam.com/top1health/healthy/20140922/20140922569504.html 中視新聞》女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 http://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="960" height="720" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> http://www.youtube.com/watch?v=j9u1lgHwXjU <iframe width="960" height="720" src="//www.youtube.com/embed/j9u1lgHwXjU" frameborder="0" allowfullscreen></iframe> cteditor cteditor 內置圖片 1 字級： 小 中 大 特 列印 轉寄 分享 太神奇！婚後產子仍是「童貞夫人」 arrow-left 1/2 arrow_right 點選放大 女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 【中視新聞】 女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 20140922 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 http://www.chinatimes.com/realtimenews/20140923001538-260405 http://www.youtube.com/watch?v=j9u1lgHwXjU <iframe width="420" height="315" src="//www.youtube.com/embed/j9u1lgHwXjU" frameborder="0" allowfullscreen></iframe> 2014年09月23日 07:44 金維中、吳烈安、曾新中 小字型 中字型 大字型 185點閱 2/10我要評比 0 影音新聞來源/中視提供 國內有超過萬名女性罹患陰道痙攣症，島至無法行房順利懷孕，嚴重一點的甚至檢查都得麻醉才能進行，更有大半數婦女因為害羞，一直不願意就診。 一名女子就是罹患重度痙攣症，結婚十幾年來從沒和丈夫行房過，但因為想要子女，醫師只能採用試管嬰兒，由於他的病情嚴重，從超音波檢查到取卵，一切都要麻醉後，才能進行。 最後女子終於順利懷的一對雙胞胎，醫師也表示，這種疾病在婦女監並不罕見，因此如果有相關病證，還是及早治療，才不會延誤到夫妻間的幸福。【中視新聞】 女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 20140922 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="480" height="360" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=blhiir8PldY <iframe width="1280" height="720" src="//www.youtube.com/embed/blhiir8PldY" frameborder="0" allowfullscreen></iframe> 收治病例的博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 -- 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四， 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 你相信嗎？1名32歲罹患嚴重陰道痙攣症的女子結婚10年，竟然丈夫從來都沒有「碰」過太太任何一次，更神奇的是，女子現已懷孕12周，將來剖腹產後仍是完璧之身，成為名副其實的「童貞夫人」！一項由國內婦產科醫師發表於台灣生殖醫學會年會上的特殊案例，令人嘖嘖稱奇。 台灣罹患陰道痙攣症(vaginismus)的婦女比例大約有1％，陰道痙攣症會嚴重影響女人進行性生活的活動無法正常進行。 婦產科醫師蔡鋒博表示，台灣罹患陰道痙攣症(vaginismus)的婦女比例大約有1％，陰道痙攣症會嚴重影響女人進行性生活的活動無法正常進行，如陰道插入：包括正常的性交，或插入衛生棉條，或使用月經杯，甚至一般婦產科醫師的內診，或使用鴨嘴做陰道檢查，都無法順利進行。 嚴重陰道痙攣症　易導致不孕 醫學上研究發現，證實患有嚴重陰道痙攣症病人，每當有外物靠近外陰部，或僅憑自己幻想可能有這樣子的動作預期，就會引起陰道肌肉群不自主強烈收縮，引發這種陰道肌肉強烈收縮的因素，包括：性交，會讓性交變的很疼痛，或根本無法正常進行，也因此使得男女無法正常性交，性伴侶無法正常在陰道射精，自然就衍生後續不孕症的問題。 這1名求診的婦女嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，所以產後仍是處女，乖可以說是難得一見的「童貞夫人」。 如果有陰道痙攣症要求子，儘早做治療要趁年輕。 陰道痙攣原因多　心理問題為主 引起陰道痙攣的原因主要與心理因素有關，多數患者由於對性知識的缺乏，對性交的極端恐懼與焦慮而產生，遭受過性暴力或外陰和陰道有創傷史的女性也易產生。以及夫妻感情不合，對配偶的不滿都有影響。 【醫師小叮嚀】： 蔡鋒博醫師表示，如果有陰道痙攣症要求子，儘早做治療要趁年輕，如果經由以下方式都失敗，包括： 1.精神科專業技巧的幫助諮詢。 2.物理治療，如由小到大的人工陰莖或鴨嘴。 3.藥物治療，如中樞神經系統肌肉鬆弛劑。 以上三個方法都失敗的話，人工助孕包括人工授精跟試管嬰兒，是重度陰道痙攣症求子的最後一張王牌！ 文章連結 http://www.top1health.com/Article/12/20293
• 2014: 陰道痙攣無性愛　卻懷了雙胞胎--- 壹週刊 nextmag: -----彰化博元婦產科蔡鋒博將在該診所因陰道痙攣進行人工生殖經驗的43例患者進行分析，發現有19個懷孕成功，其中24位人工授精病人中有8位成功懷孕，19位試管嬰兒病人中有11位懷孕成功。 ----- 彰化博元婦產科最近幫一對結婚10年無法做愛的夫妻完成人工授孕。院長蔡鋒博說，夫妻初診當天就進入試管嬰兒療程，做什麼都要麻醉，包括陰道超音波、取卵手術、植入胚胎手術、子宮鏡，還好一次就成功懷了雙胞胎，目前已12週 ------彰化博元婦產科最近幫一對從台北南下求子的陰道痙攣夫婦，9年來通通沒有性經驗，可以說是童貞夫人，麻醉後要做人工授精才發現，病人的處女膜是完整的，還好人工授精的管子非常細，就從處女膜的洞口插進去，也讓患者成功懷孕了，生完產後，處女膜還是完整。 。內置圖片 1 http://www.nextmag.com.tw/breaking-news/life/20140922/8385726 陰道痙攣無性愛卻懷了雙胞胎｜Next Magazine - 壹週刊 www.nextmag.com.tw/breaking-news/life/20140922/8385726 2 陰道痙攣是一個非常特殊又讓人難以啟齒的疾病，發生機率不到2%，這類患者無法性行為，若結了婚想生孩子怎麼辦，就得利用人工生殖來完成。 2014年09月22日 16:18 http://www.nextmag.com.tw/breaking-news/life/20140922/8385726 http://www.nextmag.com.tw/breaking-news/life/20140922/8385726 Facebook Plurk Google Plus 嚴重的陰道痙攣無法性行為，想生孩子就得靠人工授精來完成傳宗接代的任務。（資料照片） 嚴重的陰道痙攣無法性行為，想生孩子就得靠人工授精來完成傳宗接代的任務。（資料照片） 陰道痙攣是一個非常特殊又讓人難以啟齒的疾病，發生機率不到2%，這類患者無法性行為，若結了婚想生孩子怎麼辦，就得利用人工生殖來完成。彰化博元婦產科最近幫一對結婚10年無法做愛的夫妻完成人工授孕。院長蔡鋒博說，夫妻初診當天就進入試管嬰兒療程，做什麼都要麻醉，包括陰道超音波、取卵手術、植入胚胎手術、子宮鏡，還好一次就成功懷了雙胞胎，目前已12週。 日前也有一名從台北南下求子的陰道痙攣夫婦，9年來通通沒有性經驗，可以說是童貞夫人，麻醉後要做人工授精才發現，病人的處女膜是完整的，還好人工授精的管子非常細，就從處女膜的洞口插進去，也讓患者成功懷孕了，生完產後，處女膜還是完整。 蔡鋒博將在該診所因陰道痙攣進行人工生殖經驗的43例患者進行分析，發現有19個懷孕成功，其中24位人工授精病人中有8位成功懷孕，19位試管嬰兒病人中有11位懷孕成功。 蔡鋒博說，陰道痙攣在治療上，可以先進行精神科的專業諮詢；再來可由物治療來幫忙，如由小到大的人工陰莖或鴨嘴，或藉由藥物治療，如中樞神經系統肌肉鬆弛劑；若都沒效，還是無法正常行房，那麼人工生殖就是最後手段。（撰文：蔡怡真）女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四， 中視新聞》女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 國內超過萬名女性患陰道痙攣症，導致無法順利行房甚至受孕。一名三十二歲女子就因為這­樣，婚後十年從未行房、仍然是處女。因此醫師只能嘗試試管嬰兒療程，讓她順利的懷了一­對雙胞胎。 http://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="960" height="720" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> http://www.youtube.com/watch?v=j9u1lgHwXjU <iframe width="960" height="720" src="//www.youtube.com/embed/j9u1lgHwXjU" frameborder="0" allowfullscreen></iframe> 中視新聞》女陰道痙攣無法受孕 嘗試試管一舉懷雙胞 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 【中視新聞】 女陰道痙攣無法受孕 博元婦產科嘗試試管一舉懷雙胞 20140922 https://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="420" height="315" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=c_avSQZ11GA <iframe width="480" height="360" src="//www.youtube.com/embed/c_avSQZ11GA" frameborder="0" allowfullscreen></iframe> 蘋果日報報導博元婦產科的陰道痙攣症研究 中視新聞專訪報導博元婦產科蔡鋒博醫師：女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧--博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四 https://www.youtube.com/watch?v=blhiir8PldY <iframe width="1280" height="720" src="//www.youtube.com/embed/blhiir8PldY" frameborder="0" allowfullscreen></iframe> 收治病例的博元婦產科蔡鋒博醫師，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 -- 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師 女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，女陰道痙攣 終懷雙胞胎麻醉做試管助孕 剖腹產仍是完璧:婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四， 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 http://www.appledaily.com.tw/appledaily/article/headline/20140922/36100343/%E5%A5%B3%E9%99%B0%E9%81%93%E7%97%99%E6%94%A3%E7%B5%82%E6%87%B7%E9%9B%99%E8%83%9E%E8%83%8E 2014年09月22日 重度陰道痙攣症患者可藉由人工生殖方式助孕。圖為醫護人員執行人工授精作業。資料照片 【蔡明樺╱台北報導】國內逾萬名女性患陰道痙攣症，無法順利嘿咻受孕。一名三十二歲女子因重度陰道痙攣症，婚後十年從未行房、仍是處女；甚至求診時，只要器械靠近外陰部，女子就不自覺縮腿、陰道肌肉強烈收縮。為傳宗接代，嘗試試管嬰兒療程，最後一舉懷雙胞胎。醫師表示，女子現已懷孕十二周，將來剖腹產後仍是完璧之身。 收治病例的開業婦產科醫師蔡鋒博，日前在台灣生殖醫學會發表最新本土研究，統計數年來因不孕就醫的四十三名重度陰道痙攣症患者，接受人工授精、試管嬰兒療程後，十九人懷孕，成功率約四成四，證實陰道痙攣症患者想生育不再遙不可及。 別到更年期才就醫 蔡鋒博說，該名女子由丈夫陪同就醫時，主訴結婚十年仍不孕，因診間有女護理人員，女子不好啟齒原因，後來丈夫才坦承太太患陰道痙攣症，每次想嘿咻都失敗，陰莖未插入，太太就喊痛，為傳宗接代只好就醫。 蔡鋒博說，女子嘗試過心理和行為治療都依舊抗拒行房，與丈夫商量後，決定接受試管嬰兒療程。為克服她的恐懼，從陰道超音波檢查、取卵到植入胚胎等，每次都麻醉才進行，現已順利懷雙胞胎，但她不敢自然產，決定剖腹，產後仍是處女。 「陰道痙攣症治療要趁早。」高雄醫學大學婦產科教授鄭丞傑強調，很多患者年輕時就有症狀，但因怕治療，拖到更年期才就醫，連人工生殖機會都沒有。他曾收治一名四十八歲女性，結婚二十二年從沒與老公嘿咻，曾數度就醫內診，但她堅持不開腿，醫師也沒輒，拖到停經才決心想懷孕，但已來不及。 輕症者可學習放鬆 陰道痙攣症是心理影響生理，鄭丞傑臨床觀察很多患者是老師、公務員，因未有性經驗前就認為會很痛，導致親熱時不自覺縮腿、陰道肌肉緊繃。他建議症狀輕者可學習放鬆技巧，調節呼吸舒緩情緒，中重度患者可在陰道內注射肉毒桿菌素，達到放鬆肌肉效果，若都沒效則可考慮人工生殖。 陰道痙攣症小檔案 ★成因：心理影響生理，擔心第一次性行為會劇痛的想法影響所致 ★盛行率：國外研究約2％女性有此困擾，推估國內約1萬∼2萬名患者 ★症狀：害怕接受醫師內診，或行房前陰莖未插入，患者就會感到劇痛、害怕，甚至不自主雙腿夾緊、身體退縮等 ★治療： ●症狀輕微者可透過行為、心理治療，教導放鬆技巧 ●中重度患者可在陰道內注射肉毒桿菌素，舒緩肌肉緊繃 ●若上述方式仍無法自然受孕，建議採人工生殖療程助孕 資料來源：鄭丞傑醫師、蔡鋒博醫師
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導：博元婦產科找蔡鋒博醫師 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導： 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2014: 蘋果日報報導： 求子之途路遙遙 堅持15年終得三胞胎 求子歷程長達十五年的楊小姐(化名)，經歷了四次的試管嬰兒治療失敗，本來已經心灰意冷，為了不想耽誤老公的家族傳宗接代，她甚至主動跟老公提出離婚要求， 她甚至動念想主動求去 。最後在婆婆的建議下，在39歲時到博元婦產科找蔡鋒博醫師治療，想做最後一次的嘗試，沒想到竟然成功受孕，並且在母親節前夕產下三胞胎。楊小姐開心地說: 感激蔡醫師圓滿了我們的家庭! 蔡醫師特別提醒: 在試管嬰兒的治療中，”年齡”決定了成功率!台灣社會婚晚生育的狀況越來越嚴重，雖然隨著生殖醫療技術成熟，可以幫助病人解決很多”難孕”的問題。但是許多夫妻在求助於人工生殖方法時忽略了「年齡」其實才是影響懷孕的最關鍵因素。根據國民健康局統計資料顯示，到了43歲，「做人」成功率不到1成，人工懷孕率與活產率幾乎是隨著接受治療者女性的年齡之增加而下降。蔡醫師提醒想求子就要掌握35歲前的「黃金生育期」，才能提高有效受孕的成功率，也能減少在治療過程中的不適。求子之路的確十分辛苦，也充滿不確定性；也有不少病人花了許多時間在調整體質，或者使用民間偏方等方式，但卻沒有進行徹底的檢查找出真正原因，反而蹉跎時間延誤治療。依蔡醫師個人的助孕經驗，只要病人堅持，尋求正確的醫療協助，最後大有機會得到甜美的果實。堅持、努力不懈、把握時間是基本原則。
• 2013: 有沒有聽錯？53歲還能求子成功？還生雙胞胎？還一次就成功？在哪裡治療的？費用？技術？業者？ TV188
• 2013: 蝦米？卵巢腫瘤不用開刀用吸的？業者？哪裡做的？費用？要住院嗎？有傷口嗎？要麻醉嗎？手術時間？兩分鐘？ TV191
• 2013: 試管寶寶健康嗎？聰明嗎 ? 會畸形兒嗎 ? TV103
• 2013: 我年輕不懂事，墮胎太多次，導致輸卵管阻塞，幸運在博元婦產科試管 嬰兒一次成功生子 TV55
• 2013: 為何博元婦產科做試管嬰兒比較容易生龍鳳胎?秘密在? TV39
• 2013: 最新全球多囊性卵巢症候群治療最新趨勢?荷蘭共識?美國共識?大 西洋共識? TV34
• 2013: 我丈夫有精蟲抗體難怪我不能受孕，幸運的我們在博元婦產科做一次試管嬰兒就求子成功，生龍鳳胎! 精蟲抗體怎麼檢查 ? 怎麼治療 ? TV175
• 2013: 女人生小孩能力的紅、黃、綠燈?我在紅燈區?還是綠燈區?紅燈綠燈怎麼看? 如果紅燈事簿是沒救了 ? 事簿是要借卵?TV171
• 2013: 全球試管嬰兒最新科技，最新儀器大公開!新的儀器新的科技可大幅提昇試管嬰兒懷孕率 TV172
• 2013: 我卵巢早衰，我在博元婦產科借卵生子成功 TV173
• 2013: 我無精，我在博元婦產科借精生子成功 TV174
• 2013: 我兒子有精子，我要他借精生子!為什麼有精要借精?有什麼疾病? 有什麼隱情 ?TV174
• 2013: 53歲在博元婦產科求子成功 ?為什麼53歲還要生孩子? 博元婦產科有什麼新技術 ?TV188
• 2013: 我年輕不懂事，墮胎太多次，婚後不孕，博元蔡醫師用子宮鏡檢查 說我有子宮沾黏息肉!經子宮鏡治療我自然懷孕了 TV109
• 2013: 我不想做試管嬰兒，我要做什麼可增加自然受孕 的機會!TV100
• 2013: 我的精索靜脈曲張，睪丸痛，精子少，不孕，要怎麼治療才會懷孕？！博元 婦產科很專業! TV106
• 2013: 胚胎植入前基因診斷（PGD） 可提高試管嬰兒成功率！哪裡有？費用？很貴嗎？怎麼做？ TV23
• 2013: 我子宮感染性病，常常白帶，陰道發炎，披衣菌，梅毒，淋病，會不孕症嗎 TV125
• 2013: 奇怪？為何我的胚胎不著床？有什麼新科技可以增加胚胎著床率？有什麼新科技可以增加懷孕率？ TV101
• 2013: 蝦米？不到30歲就停經？什麼是卵巢未老先衰？怎麼治療才能夠生小孩？博元婦產科 TV99
• 2013: 26秒用吸的(不是用刮的)中止妊娠手術(即人工流產)，不會不孕，不痛，安全！照片由上到下就是手術過程
• 2013: 為什麼幹細胞可以終結不孕？什麼是人造精子？人造卵子？ TV96
• 2013: 吃什麼容易受孕？山珍海味？雞精？山藥？中藥？蜂王乳？ TV104
• 2013: 我在博元婦產科求子成功了：丈夫少精症 ，多少精子才算少？多少精子才需做試管嬰兒？TV114
• 2013: 女人生孩子能力的紅燈、黃燈、綠燈，妳是什麼燈？紅燈？危險嗎？有救嗎？ TV171
• 2013: 林青蓉追問博元婦產科蔡鋒博醫師 ：什麼是第三代基因晶片試管嬰兒 ？
• 2013: 我卵巢功能不佳，如何求子才能容易懷孕？博元婦產科 TV112
• 2013: 什麼是精蟲抗體？怎麼檢查？我在博元婦產科求子成功 TV175
• 2013: 夫妻都有遺傳疾病，如海洋性貧血，要如何避免生下遺傳疾病海 洋性貧血小孩？胚胎切片， 第三代試管嬰兒，基因晶片 TV117
• 2013: 向博元婦產科卵子銀行借卵需要費用嗎？怎麼借？程序？ TV168
• 2013: 做完子宮鏡，就懷孕！當真？ TV189
• 2013: 我在博元婦產科只做一次人工授精，求子成功 TV179
• 2013: 久久神功也會不孕！射精異常也會不孕！在博元婦產科求子成功 TV193
• 2013: 我沒子宮想要找代理孕母，怎麼代？怎麼找？合法嗎？會通過立法嗎？ TV197
• 2013: 迷你試管嬰兒，多迷你？費用迷你嗎？打針迷你嗎？不必打針嗎？少打很多針嗎？ TV199
• 2013: 天啊！蔡醫師做個子宮鏡，竟發現她的子宮蓋了樓中樓，經蔡醫師手術後做試管嬰兒，成功生下一男嬰！ TV189
• 2013: 蝦米？夫妻八字不合也會不孕？談抗精蟲抗體引起的不孕，怎麼檢 查？到哪裡檢查？費用？怎麼治療才能夠懷孕？（上） TV14
• 2013: 蝦米？殺手細胞殺胚胎導致流產？子宮內膜殺手細胞過多引起的 習慣性流產？到哪裡檢查？博元婦產科！費用？ 怎麼治療？TV29
• 2013: 一對不孕症夫妻要做哪些檢查，才能找出不孕症的原因？才能快速懷孕？ TV2
• 2013: 我在博元婦產科做一次試管嬰兒就成功啦！她和博元試管寶寶上電視臺現 身說法，分享她的成功求子經驗 TV187
• 2013: 真人真事：我在博元婦產科做一次試管嬰兒就成功啦！她和博元試 管寶寶上電視臺現 身說法，分享她的成功求子經驗 TV187
• 2013: 逆行性射精？糖尿病精子跑到尿液裡面去？怎麼生小孩？大海撈精，博元試管嬰兒成功幫助病人懷孕生子案例分析： TV42
• 2013: 蝦米？做一次人工授精就生雙胞胎， 成功案例分析：TV40
• 2013: 什麼是胚胎強力膠？聽說可增加試管嬰兒成功率！哪裡有？費用？ TV30
• 2013: 不孕四年，輸卵管不通，卵巢功能不佳AMH=0.65 ，由慈O醫院轉診過來在博元婦產科只做一次試管嬰兒，陳〇〇有了！"
• 2013: 氣死我了，每次懷孕就流產，流流流，流到都沒有生孩子的信心 了，談習慣性流產怎麼治療？博元婦產科 TV27
• 2013: 如何使53歲，都已經停經4年，在博 元婦產科只做一次試管嬰兒雙胞胎 ？請聽蔡醫師分析如何辦 到？ 博元試管嬰兒成功案例分析： TV36
• 2013: 全世界多囊性卵巢症候群治療最新趨勢，博元婦產科分析給你聽， 報乎你知 TV34
• 2013: 不孕症4年 AMH只有0.65， 輸卵管阻塞, 在博元婦產科做一次試管嬰兒，就成功懷孕
• 2013: 在胚胎的殼用雷射打個洞這樣可以增加試管嬰兒成功率？哪 裡有？博元婦產科！費用？安全嗎？危險嗎？雷 射胚胎輔助孵化 TV81
• 2013: 我需要做人工授精嗎？費用？怎麼做？哪裡做？博元婦產科！ 哪些人要做人工授精 TV8
• 2013: 博元試管嬰兒太多的成功案例：非常多求子趣談，和你分享，他們 可以做到、你也可以做到 TV66
• 2013: ​雙角子宮怎麼懷孕？雙子宮怎麼懷孕？子宮有隔間怎麼懷孕？博元婦產科​搶救子宮 ​，助妳懷孕​ TV202
• 2013: 我有多曩性卵巢，我在博元婦產科做試管嬰兒成功懷孕！　TV111
• 2013: 27歲就停經？卵巢未老先衰？怎麼懷 孕？借卵？我不要！有藥醫 嗎？ TV99
• 2013: 為什麼用超音波，植胚胎可提高試管嬰兒成功懷孕率？ TV190
• 2013: 博元婦產科詳細介紹試管嬰兒新科技 TV172
• 2013: 試管嬰兒新科技可提升試管嬰兒成功率， 由博元婦產科詳細一一介紹 TV172
• 2013: (尚未設定標題)
• 2013: 林杰樑最後研究 鉛殺精蟲 害不孕
• 2013: 力不從心 遇上狼虎之年…
• 2013: 蝦米？射精射到膀胱？射精異常怎麼懷孕？博元婦產科助妳求子成功 TV193
• 2013: 試管嬰兒怎麼做才能懷孕成功？博元婦產科告訴妳 TV9
• 2013: 什麼是胚胎強力膠？哪裡有？博元婦產科！費用？為什麼可以增加試管嬰兒成功率？ TV30
• 2013: ​哪裡有在檢查​免疫引起的習慣性流產​？檢查什麼？抽 ​寫嗎？子宮鏡嗎？殺手細胞嗎？​ 博元婦產科​ TV29
• 2013: 量身訂做的試管嬰兒？怎麼量身訂做？費用？怎麼做？哪裡有做？ TV47
• 2013: ​子宮頸閉鎖不全，​子宮頸 ​引起的​不孕​症，有哪一些人？怎麼治療才能夠成功懷孕 ​？​ TV52
• 2013: 她借卵子成功生子了！在哪裡借的？博元婦產科 TV54
• 2013: 女人卵巢功能怎麼看好壞？到哪裡檢查？費用？項目？什麼時候 檢查？ TV57
• 2013: 怎麼準備？試管嬰兒才容易一次成功？ TV160