Objective: To evaluate the effect of preimplantation genetic diagnosis (PGD) conducted for women who had Down syndrome pregnancy previously. Methods: Trisomy 21 was diagnosed by using fluorescence in site hybridizatio...Objective: To evaluate the effect of preimplantation genetic diagnosis (PGD) conducted for women who had Down syndrome pregnancy previously. Methods: Trisomy 21 was diagnosed by using fluorescence in site hybridization (FISH) before embryo transfer in two women who had Down syndrome pregnancies. Each received one or two PGD cycles respectively. Results: Case 1: one PGD cycle was conducted, two oocytes were fertilized and biopsied. One embryo is of trisomy 21 and the other of monosomy 21. No embryo was transferred. Case 2: two PGD cycles were conducted, in total, sixteen oocytes were fertilized and biopsied. Four embryos were tested to be normal, six of trisomy 21, and one of monosomy 21. Five had no signal. Four normal embryos were transferred but no pregnancy resulted. Conclusion: For couples who had pregnancies with Down syndrome pre-viously, PGD can be considered, and has been shown to be an effective strategy.展开更多
Preimplantation genetic diagnosis (PGD), as a new assisted reproductive technology which can select normal embryos for transplantation combined with in-vitro fertilization and embryo transfer(IVF-ET)through the analys...Preimplantation genetic diagnosis (PGD), as a new assisted reproductive technology which can select normal embryos for transplantation combined with in-vitro fertilization and embryo transfer(IVF-ET)through the analysis of genetic materials before embryo implantation, holds a more and more important position in the diagnosis of genetic diseases and has made an important significance to the Aristogenics.PGD is an important aspect of assisted reproduction technology(ART)with its rapid development. Continuous appearances and comprehensive applications of new methods and technologies have greatly developed the PGD. In this review, we introduce some new methods and their principles about the new research advances of PGD.展开更多
Reliable and accurate pre-implantation genetic diagnosis （PGD） of patient＇s embryos by next-generation sequencing （NGS） is dependent on efficient whole genome amplification （WGA） of a representative biopsy samp...Reliable and accurate pre-implantation genetic diagnosis （PGD） of patient＇s embryos by next-generation sequencing （NGS） is dependent on efficient whole genome amplification （WGA） of a representative biopsy sample. However, the performance of the current state of the art WGA methods has not been evaluated for sequencing. Using low template DNA （15 pg） and single cells, we showed that the two PCR-based WGA systems SurePlex and MALBAC are superior to the REPLI-g WGA multiple displacement amplification （MDA） system in terms of consistent and reproducible genome coverage and sequence bias across the 24 chromosomes, allowing better normalization of test to reference sequencing data. When copy number variation sequencing （CNV-Seq） was applied to single cell WGA products derived by either SurePlex or MALBAC amplification, we showed that known disease CNVs in the range of 3-15 Mb could be reliably and accurately detected at the correct genomic positions. These findings indicate that our CNV-Seq pipeline incorporating either SurePlex or MALBAC as the key initial WGA step is a powerful methodology for clinical PGD to identify euploid embryos in a patient＇s cohort for uterine transplantation,展开更多
Objective To investigate the feasibility of vitrification of blastocysts following blastomere biopsy. Methods Among patients undergoing pre-implantation genetic diagnosis （PGD）, artificial shrinkage of the blastocoe...Objective To investigate the feasibility of vitrification of blastocysts following blastomere biopsy. Methods Among patients undergoing pre-implantation genetic diagnosis （PGD）, artificial shrinkage of the blastocoelic cavity and subsequent vitrification of applicable surplus blastocysts after day-3 blastomere biopsy were performed. According to patient requirements, thawed blastocysts were transferred into patients due to pregnancy failure after fresh embryo transfer, ectopic pregnancy, ovarian hyperstimulation. Results Twenty-four PGD cycles were carried out. According to genetic diagnosis and the development of blastocysts, transfer was cancelled in 7 cycles due to absence of applicable embryos or ovarian hyperstimulation. In the remaining 17 cycles, 26 blastocysts were thawed and transferred, which resulted in 13 implanted （50.0%）. Clinical pregnancies were observed in 11 patients （64.71%）. Following transfer, 30 applicable blastocysts in 10 cycles were cryopreserved. Six patients received transfer of thawed blastocysts. All 8 thawed embryos survived and were transferred, and singleton pregnancies occurred in 5 patients. Two women delivered healthy infants and 3 pregnancies are ongoing. Conclusion Vitrification with artificial shrinkage is effective for preserving blastocysts following blastomere biopsy.展开更多
目的探讨高孕激素下促排卵方案和黄体期长方案对平衡易位携带行PGD周期治疗效果的比较。方法回顾性分析2016年5月至2018年4月在武汉大学人民医院生殖医学中心因染色体平衡易位行PGD助孕的51例患者的临床资料,根据患者促排卵方案的不同...目的探讨高孕激素下促排卵方案和黄体期长方案对平衡易位携带行PGD周期治疗效果的比较。方法回顾性分析2016年5月至2018年4月在武汉大学人民医院生殖医学中心因染色体平衡易位行PGD助孕的51例患者的临床资料,根据患者促排卵方案的不同将其分为高孕激素下促排卵方案组(PPOS方案组)和黄体期长方案组(LP方案组),其中PPOS方案组24例患者,LP方案组27例患者。比较两组患者的一般特点、促排卵情况、胚胎发育情况和子代胚胎染色体情况。结果两组患者的一般情况,包括双方年龄、不孕年限、BMI、基础窦卵泡数、基础FSH、基础LH、FSH/LH比值和基础E2比较,其差异均无统计学意义(均P>0.05)。促排卵过程中,PPOS方案组患者促排天数显著低于LP方案组患者(9.21±1.67 vs 11.22±2.87,P=0.003),PPOS方案组患者获卵数少于LP方案组患者[10.50(8.25,17.50)vs 14.00(12.00,20.00),P=0.096)],PPOS方案组患者M2卵子数少于LP方案组患者[9.50(7.00,13.00) vs 12.00(8.00,18.00),P=0.219)],PPOS方案组患者2PN数少于LP方案组患者[8.00(6.00,12.00) vs 9.00(5.00,14.00),P=0.583)],PPOS方案组患者第5天囊胚比例高于LP方案组患者(56.12%vs 45.65%,P=0.149),两组差异均无统计学意义;但PPOS方案组患者可活检囊胚形成率显著高于LP方案组患者(44.34%vs 34.40%,P=0.023),其差异具有统计学意义。两组患者胚胎染色体形成情况中,正常/平衡胚胎比例(32.65%vs 36.08%,P=0.614))、不平衡胚胎比例(40.82%vs 32.99%,P=0.495))、非整倍体胚胎比例(37.76%vs 41.24%,P=0.619))等比较,其差异均无统计学意义。2种方案均有约30%的患者无可移植胚胎(29.17%vs 29.63%,P=0.971)。结论对于染色体平衡易位行PGD助孕的患者,PPOS方案能够获得与黄体期长方案相似的促排卵效果,而且可活检囊胚形成率更高。通过分析二代测序结果显示,PPOS方案对子代染色体形成没有不利影响。所以,对于染色体平衡易位行PGD助孕的患者,PPOS方案是一种更为经济合理的促排卵方案。展开更多
Birth defects are caused by multiple factors,such as chromosome abnormality,environmental factors,and maternal factors.In this study,we focused on exploring the genetic causes of a non-consanguineous couple who suffer...Birth defects are caused by multiple factors,such as chromosome abnormality,environmental factors,and maternal factors.In this study,we focused on exploring the genetic causes of a non-consanguineous couple who suffered from four times of unsuccessful pregnancy due to unexplained recurrent fetal malformations with similar symptoms and normal chromosome copy number variations.Using trio-whole exome sequencing(trio-WES) for this couple and one of the affected fetuses,we found a mutation,c.1996 delC on the maternal imprinted gene MAGEL2 that was carried by the affected fetus and husband,leading to Schaaf-Yang syndrome.To screen this mutation,we further performed preimplantation genetic diagnosis(PGD) strategy followed by a gene pedigree validation and pathogenicity analysis.After the transfer of a PGD-screened embryo,a normal newborn without previous abnormal symptoms was born(February 15,2019).We present the first data that identified a pathogenic gene(MAGEL2 c.1996 delC) in a fetus with Schaaf-Yang syndrome in the EAS(East Asian) database and overcame this genetic defect by using processed PGD for this couple based on the WES results.展开更多
A couple with a proband child of GJB2 （encoding the gap junction protein connexin 26）-associated hearing impairment and a previous pregnancy miscarriage sought for a reproductive solution to bear a healthy child. Ou...A couple with a proband child of GJB2 （encoding the gap junction protein connexin 26）-associated hearing impairment and a previous pregnancy miscarriage sought for a reproductive solution to bear a healthy child. Our study aimed to develop a cus- tomized preconception-to-neonate care trajectory to fulfill this clinical demand by integrating preimplantation genetic diagno- sis （PGD）, noninvasive prenatal testing （NIPT）, and noninvasive prenatal diagnosis （N1PD） into the strategy. Auditory and ge- netic diagnosis of the proband child was carried out to identify the disease causative mutations. The couple then received in-vitro-fertilization treatment, and eight embryos were obtained for day 5 biopsy. PGD was performed by short-tandem-repeat linkage analysis and Sanger sequencing of GJB2 gene. Transfer of a GJB2c.235delC heterozygous embryo resulted in a sin- gleton pregnancy. At the 13th week of gestation, genomic DNA （gDNA） from the trio family and cell-free DNA （cfDNA） from maternal plasma were obtained for assessment of fetal chromosomal aneuploidy and GJB2 mutations. NIPT and NIPD showed the absence of chromosomal aneuploidy and GJB2-associated disease in the fetus, which was later confirmed by inva- sire procedures and postnatal genetic/auditory diagnosis. This strategy successfully prevented the transmission of hearing im- pairment in the newborn, thus providing a valuable experience in reproductive management of similar cases and potentially other monogenic disorders.展开更多
植入前遗传学诊断(Preimplantation Genetic D iagnosis PGD)是辅助生育技术与分子生物学技术相结合而发展的孕前遗传学诊断技术,在植入子宫前淘汰了遗传异常的胚胎,是产前诊断技术的重大进展。但是,由于技术本身存在着一定局限性和不...植入前遗传学诊断(Preimplantation Genetic D iagnosis PGD)是辅助生育技术与分子生物学技术相结合而发展的孕前遗传学诊断技术,在植入子宫前淘汰了遗传异常的胚胎,是产前诊断技术的重大进展。但是,由于技术本身存在着一定局限性和不确定性,同时,受到病人认知能力等因素的影响,由此引发了系列伦理学争议。在进行PGD前,一个明了、详尽的患者知情同意过程是必须的。包括通俗全面告知PGD有关信息、手术和检测的局限性和可能结果;充分告知通过PGD所获得的利益和风险。在此基础上针对不同的遗传病检测签署详细的书面知情同意书。展开更多
基金Project supported by the National Basic Research Program of China(Nos. 2006CB944006 and 2006CB504004)the Key Research Pro-gram of Zhejiang Province, China (No. 2006C13078)the Bureau of Science and Technology of Hangzhou, China (No. 20061123B03)
文摘Objective: To evaluate the effect of preimplantation genetic diagnosis (PGD) conducted for women who had Down syndrome pregnancy previously. Methods: Trisomy 21 was diagnosed by using fluorescence in site hybridization (FISH) before embryo transfer in two women who had Down syndrome pregnancies. Each received one or two PGD cycles respectively. Results: Case 1: one PGD cycle was conducted, two oocytes were fertilized and biopsied. One embryo is of trisomy 21 and the other of monosomy 21. No embryo was transferred. Case 2: two PGD cycles were conducted, in total, sixteen oocytes were fertilized and biopsied. Four embryos were tested to be normal, six of trisomy 21, and one of monosomy 21. Five had no signal. Four normal embryos were transferred but no pregnancy resulted. Conclusion: For couples who had pregnancies with Down syndrome pre-viously, PGD can be considered, and has been shown to be an effective strategy.
基金National Natural Science Foundation of China ( 30772069)
文摘Preimplantation genetic diagnosis (PGD), as a new assisted reproductive technology which can select normal embryos for transplantation combined with in-vitro fertilization and embryo transfer(IVF-ET)through the analysis of genetic materials before embryo implantation, holds a more and more important position in the diagnosis of genetic diseases and has made an important significance to the Aristogenics.PGD is an important aspect of assisted reproduction technology(ART)with its rapid development. Continuous appearances and comprehensive applications of new methods and technologies have greatly developed the PGD. In this review, we introduce some new methods and their principles about the new research advances of PGD.
基金supported by grants awarded to Yuanqing Yao by the Key Program of the "Twelfth Five-year plan" of People’s liberation Army(No.BWS11J058)the National High Technology Research and Development Program(SS2015AA020402)
文摘Reliable and accurate pre-implantation genetic diagnosis （PGD） of patient＇s embryos by next-generation sequencing （NGS） is dependent on efficient whole genome amplification （WGA） of a representative biopsy sample. However, the performance of the current state of the art WGA methods has not been evaluated for sequencing. Using low template DNA （15 pg） and single cells, we showed that the two PCR-based WGA systems SurePlex and MALBAC are superior to the REPLI-g WGA multiple displacement amplification （MDA） system in terms of consistent and reproducible genome coverage and sequence bias across the 24 chromosomes, allowing better normalization of test to reference sequencing data. When copy number variation sequencing （CNV-Seq） was applied to single cell WGA products derived by either SurePlex or MALBAC amplification, we showed that known disease CNVs in the range of 3-15 Mb could be reliably and accurately detected at the correct genomic positions. These findings indicate that our CNV-Seq pipeline incorporating either SurePlex or MALBAC as the key initial WGA step is a powerful methodology for clinical PGD to identify euploid embryos in a patient＇s cohort for uterine transplantation,
基金funded by Guangxi Zhuang Autonomous Region Natural Science Foundation of China (Grant No. 0897007, 0832183, 0542058)Health Department of Guangxi Zhuang Autonomous Region (Grant No. 200947, Z2007013)
文摘Objective To investigate the feasibility of vitrification of blastocysts following blastomere biopsy. Methods Among patients undergoing pre-implantation genetic diagnosis （PGD）, artificial shrinkage of the blastocoelic cavity and subsequent vitrification of applicable surplus blastocysts after day-3 blastomere biopsy were performed. According to patient requirements, thawed blastocysts were transferred into patients due to pregnancy failure after fresh embryo transfer, ectopic pregnancy, ovarian hyperstimulation. Results Twenty-four PGD cycles were carried out. According to genetic diagnosis and the development of blastocysts, transfer was cancelled in 7 cycles due to absence of applicable embryos or ovarian hyperstimulation. In the remaining 17 cycles, 26 blastocysts were thawed and transferred, which resulted in 13 implanted （50.0%）. Clinical pregnancies were observed in 11 patients （64.71%）. Following transfer, 30 applicable blastocysts in 10 cycles were cryopreserved. Six patients received transfer of thawed blastocysts. All 8 thawed embryos survived and were transferred, and singleton pregnancies occurred in 5 patients. Two women delivered healthy infants and 3 pregnancies are ongoing. Conclusion Vitrification with artificial shrinkage is effective for preserving blastocysts following blastomere biopsy.
文摘目的探讨高孕激素下促排卵方案和黄体期长方案对平衡易位携带行PGD周期治疗效果的比较。方法回顾性分析2016年5月至2018年4月在武汉大学人民医院生殖医学中心因染色体平衡易位行PGD助孕的51例患者的临床资料,根据患者促排卵方案的不同将其分为高孕激素下促排卵方案组(PPOS方案组)和黄体期长方案组(LP方案组),其中PPOS方案组24例患者,LP方案组27例患者。比较两组患者的一般特点、促排卵情况、胚胎发育情况和子代胚胎染色体情况。结果两组患者的一般情况,包括双方年龄、不孕年限、BMI、基础窦卵泡数、基础FSH、基础LH、FSH/LH比值和基础E2比较,其差异均无统计学意义(均P>0.05)。促排卵过程中,PPOS方案组患者促排天数显著低于LP方案组患者(9.21±1.67 vs 11.22±2.87,P=0.003),PPOS方案组患者获卵数少于LP方案组患者[10.50(8.25,17.50)vs 14.00(12.00,20.00),P=0.096)],PPOS方案组患者M2卵子数少于LP方案组患者[9.50(7.00,13.00) vs 12.00(8.00,18.00),P=0.219)],PPOS方案组患者2PN数少于LP方案组患者[8.00(6.00,12.00) vs 9.00(5.00,14.00),P=0.583)],PPOS方案组患者第5天囊胚比例高于LP方案组患者(56.12%vs 45.65%,P=0.149),两组差异均无统计学意义;但PPOS方案组患者可活检囊胚形成率显著高于LP方案组患者(44.34%vs 34.40%,P=0.023),其差异具有统计学意义。两组患者胚胎染色体形成情况中,正常/平衡胚胎比例(32.65%vs 36.08%,P=0.614))、不平衡胚胎比例(40.82%vs 32.99%,P=0.495))、非整倍体胚胎比例(37.76%vs 41.24%,P=0.619))等比较,其差异均无统计学意义。2种方案均有约30%的患者无可移植胚胎(29.17%vs 29.63%,P=0.971)。结论对于染色体平衡易位行PGD助孕的患者,PPOS方案能够获得与黄体期长方案相似的促排卵效果,而且可活检囊胚形成率更高。通过分析二代测序结果显示,PPOS方案对子代染色体形成没有不利影响。所以,对于染色体平衡易位行PGD助孕的患者,PPOS方案是一种更为经济合理的促排卵方案。
基金supported by the National Natural Science Foundation of China(31522034,81521002,81730038)the National High Technology Research and Development Program(2015AA020407)
文摘Birth defects are caused by multiple factors,such as chromosome abnormality,environmental factors,and maternal factors.In this study,we focused on exploring the genetic causes of a non-consanguineous couple who suffered from four times of unsuccessful pregnancy due to unexplained recurrent fetal malformations with similar symptoms and normal chromosome copy number variations.Using trio-whole exome sequencing(trio-WES) for this couple and one of the affected fetuses,we found a mutation,c.1996 delC on the maternal imprinted gene MAGEL2 that was carried by the affected fetus and husband,leading to Schaaf-Yang syndrome.To screen this mutation,we further performed preimplantation genetic diagnosis(PGD) strategy followed by a gene pedigree validation and pathogenicity analysis.After the transfer of a PGD-screened embryo,a normal newborn without previous abnormal symptoms was born(February 15,2019).We present the first data that identified a pathogenic gene(MAGEL2 c.1996 delC) in a fetus with Schaaf-Yang syndrome in the EAS(East Asian) database and overcame this genetic defect by using processed PGD for this couple based on the WES results.
基金supported by the National Program on Key Basic Research Project(2014CB943001 and 2012CB944700)the National Natural Science Foundation of China(81120108009 and 81530032)+3 种基金the National Health and Family Planning Commission of the People's Republic of China(201402004)Science and Technology Plan of Guangdong Province(2013B022000005)Guangdong Enterprise Key Laboratory of Human Disease Genomics(2011A060906007)Shenzhen Engineering Laboratory for Birth Defects Screening([2011]861)
文摘A couple with a proband child of GJB2 （encoding the gap junction protein connexin 26）-associated hearing impairment and a previous pregnancy miscarriage sought for a reproductive solution to bear a healthy child. Our study aimed to develop a cus- tomized preconception-to-neonate care trajectory to fulfill this clinical demand by integrating preimplantation genetic diagno- sis （PGD）, noninvasive prenatal testing （NIPT）, and noninvasive prenatal diagnosis （N1PD） into the strategy. Auditory and ge- netic diagnosis of the proband child was carried out to identify the disease causative mutations. The couple then received in-vitro-fertilization treatment, and eight embryos were obtained for day 5 biopsy. PGD was performed by short-tandem-repeat linkage analysis and Sanger sequencing of GJB2 gene. Transfer of a GJB2c.235delC heterozygous embryo resulted in a sin- gleton pregnancy. At the 13th week of gestation, genomic DNA （gDNA） from the trio family and cell-free DNA （cfDNA） from maternal plasma were obtained for assessment of fetal chromosomal aneuploidy and GJB2 mutations. NIPT and NIPD showed the absence of chromosomal aneuploidy and GJB2-associated disease in the fetus, which was later confirmed by inva- sire procedures and postnatal genetic/auditory diagnosis. This strategy successfully prevented the transmission of hearing im- pairment in the newborn, thus providing a valuable experience in reproductive management of similar cases and potentially other monogenic disorders.
文摘植入前遗传学诊断(Preimplantation Genetic D iagnosis PGD)是辅助生育技术与分子生物学技术相结合而发展的孕前遗传学诊断技术,在植入子宫前淘汰了遗传异常的胚胎,是产前诊断技术的重大进展。但是,由于技术本身存在着一定局限性和不确定性,同时,受到病人认知能力等因素的影响,由此引发了系列伦理学争议。在进行PGD前,一个明了、详尽的患者知情同意过程是必须的。包括通俗全面告知PGD有关信息、手术和检测的局限性和可能结果;充分告知通过PGD所获得的利益和风险。在此基础上针对不同的遗传病检测签署详细的书面知情同意书。
