Preimplantation genetic testing for aneuploidy improves clinical outcomes in patients with repeated implantation failure

Objective:The objective of this study is to study whether preimplantation genetic testing for aneuploidy(PGT-A)improves the clinical outcomes of infertile patients with repeated implantation failure(RIF)undergoing frozen-thawed embryo transfer.Methods:This is a retrospective analysis of clinical pregnancy,live birth,miscarriage rates,and obstetric and perinatal outcomes of women with RIF with or without PGT-A.Statistical analyses of categorical data were performed using propensity score matching(PSM),χ^(2)test,and Student’s t test.Results:We enrolled 466 patients with RIF,of which,209 were in the RIF-PGT-A group.The rate of euploid blastocysts was significantly associated with age and day 5 or 6 blastocysts.There were significant differences between the RIF-PGT-A group and the RIF-non-PGT-A group across several parameters.After PSM,positive serum human chorionic gonadotropin(56.9%and 33.9%,P<0.01),clinical pregnancy(49.5%and 31.2%,P<0.01),live birth(43.1%and 25.7%,P<0.01),and fetal heart rates(50.0%and 29.8%,P<0.01)per transfer were significantly higher in the RIF-PGT-A group.Conclusion:Elective single-embryo transfer PGT-A can minimize the risk of obstetric and perinatal outcomes,especially fetal body weight,in women with RIF.Additionally,PGT-A can significantly improve pregnancy and live birth rates.

Preimplantation HLA typing: Practical tool for stem cell transplantation treatment of congenital disorders

It is well known that to achieve an acceptable engraftment and survival in stem cell therapy, an human leukocyte antigens(HLA) identical stem cell transplant is strongly required. However, the availability of the HLA matched donors even among family members is extremely limited, so preimplantation HLA typing provides an attractive practical tool of stem cell therapy for children requiring HLA matched stem cell transplantation. The present experience of preimplantation genetic diagnosis(PGD) for HLA typing of over one thousand cases shows that PGD provides the at-risk couples with the option to establish an unaffected pregnancy, which may benefit the affected member of the family with hemoglobinopathies, immunodeficiencies and other congenital or acquired bone marrow failures. Despite ethical issues involved in preimplantation HLA typing, the data presented below show an extremely high attractiveness of this option for the couples with affected children requiring HLA compatible stem cell transplantation.

低分子肝素及胚胎植入前非整倍体检测对不明原因复发性流产患者妊娠结局的影响

目的:比较行胚胎植入前非整倍体检测(PGT-A)助孕和未行PGT-A助孕的不明原因复发性流产(URSA)患者应用低分子肝素(LMWH)后临床结局的差异。方法:回顾分析2016年1月1日至2022年5月17日URSA患者于郑州大学第三附属医院生殖医学科首次移植冷冻囊胚共1158个周期的临床资料,按移植前是否行PGT-A分为PGT-A组(行PGT-A+ICSI,315例)和非PGT-A组(行IVF/ICSI,843例),再根据移植日是否使用LMWH分为肝素组和非肝素组,为调整混杂因素进行1∶5倾向性评分匹配(PSM),对PSM后患者数据进行分析,比较两组患者种植率、临床妊娠率、流产率、早期流产率、活产率及单胎活产围产结局的差异。对非PGT-A组和PGT-A组PSM前数据、肝素PGT-A组和肝素非PGT-A组临床妊娠结局进行多因素logistic回归分析。结果:非PGT-A组中,肝素组和非肝素组的不孕年限差异有统计学意义(P<0.05),种植率(33.8%vs 32.1%,P=0.610)、临床妊娠率(45.4%vs 43.8%,P=0.733)、流产率(28.0%vs 28.4%,P=0.958)、早期流产率(23.8%vs 25.4%,P=0.797)、活产率(31.9%vs 30.1%,P=0.671)比较差异无统计学意义(P>0.05)。PGT-A组中,肝素组和非肝素组的女方体质量指数(BMI)比较差异有统计学意义(P<0.05),种植率(59.8%vs 57.6%,P=0.763)、临床妊娠率(59.4%vs 57.6%,P=0.806)、流产率(20.4%vs 14.7%,P=0.448)、早期流产率(16.4%vs 14.7%,P=0.803)、活产率(46.9%vs 49.2%,P=0.752)比较差异无统计学意义(P>0.05)。非PGT-A组中,肝素组和非肝素组内获得单胎活产患者的围产期结局比较差异均无统计学意义(P>0.05);PGT-A组中,肝素组较非肝素组小于胎龄儿(10.3%vs 0.8%,P=0.023),差异有统计学意义(P<0.05),余获得单胎活产患者的围产期结局差异均无统计学意义(P>0.05)。结论:行PGT-A助孕和未行PGT-A助孕的URSA患者应用LMWH并不能改善其妊娠结局,且行PGT-A助孕应用LMWH患者的围产期小于胎龄儿发生风险显著升高。

Retrospective Cohort Study of Preimplantation Genetic Testing for Aneuploidy with Comprehensive Chromosome Screening versus Nonpreimplantation Genetic Testing in Normal Karyotype,Secondary Infertility Patients with Recurrent Pregnancy Loss

Objective:To evaluate whether preimplantation genetic testing for aneuploidy(PGT-A)with comprehensive chromosome screening increases live birth rate(LBR)in normal karyotype couples with recurrent pregnancy loss(RPL).Methods:A retrospective cohort follow-up study of 506 couples with RPL was conducted between April 2014 and March 2017.Couples were allocated to two groups according to their decision to choose PGT-A or not.The primary outcome was LBR per start/transfer cycle;secondary outcomes were ongoing pregnancy rate and miscarriage rate.Statistical analyses were conducted using univariate and multivariate logistic regression models adjusted for maternal age.Results:LBR per start(26.6%vs.15.4%,relative risk[RR]:2.66,95%confidence interval[CI][1.69-4.20],P<0.0001;adjusted RR[aRR]:2.40,95%CI[1.49-3.86],P=0.0004)and per transfer(44.9%vs.25.1%,RR:3.00,95%CI[1.96-4.60],P<0.0001;aRR:2.64,95%CI[1.68-4.14],P<0.0001)was significantly higher in the PGT-A group than in the non-PGT-A group.The miscarriage rate was significantly lower in the PGT-A group compared to the non-PGT-A group(15.7%vs.34.6%,RR:0.27,95%CI[0.13-0.57],P=0.00005;aRR:0.26,95%CI[0.12-0.57],P=0.0007).Conclusions:LBR per start cycle following PGT-A is significantly higher,and risk of miscarriage is significantly lower among infertile couples with RPL,irrespective of maternal age.PGT-A should be recommended to infertile couples with RPL.

In vitro Fertilization with Single-nucleotide Polymorphism Microarray-based Preimplantation Genetic Testing for Aneuploidy Significantly Improves Clinical Outcomes in Infertile Women with Recurrent Pregnancy Loss:A Randomized Controlled Trial

Objective:To evaluate the effect of preimplantation genetic testing for aneuploidy(PGT-A)in infertile patients with recurrent pregnancy loss(RPL).Methods:A prospective randomized clinical trial was performed in a university-affiliated fertility center in Shanghai,China.Patients in the PGT-A group underwent blastocyst biopsy followed by single-nucleotide polymorphism microarray-based PGT-A and single euploid blastocyst transfer,whereas patients in the control group underwent routine in vitro fertilization/ICSI procedures and frozen embryo transfer of 1-2 embryos selected according to morphological standards.Results:Two hundred and seven infertile patients with RPL were included in this study and randomly assigned to either the control or the PGT-A group.Baseline variables and cycle characteristics were comparable between the two groups.The results showed that PGT-A significantly improved the ongoing pregnancy rate(55.34%vs.29.81%)as well as the live birth rate(48.54%vs.27.88%)and significantly reduced the miscarriage rate(0.00%vs.14.42%)on a per-patient analysis.A significant increase in cumulative ongoing pregnancy rates over time was observed in the PGT-A group.Subgroup analysis showed that the significant benefit diminished for patients who attempted≥2 PGT-A cycles.Conclusions:PGT-A significantly improved the ongoing pregnancy and live birth rate,while reduced miscarriage rate in infertile RPL patients.However,the significance diminished in patients attempting≥2 cycles;thus,further studies are warranted to explore the most cost-effective number of attempts in these patients to avoid overuse.

生长激素预处理在胚胎植入前染色体非整倍体检测中的应用研究

目的研究生长激素(GH)预处理对整倍体的改善及妊娠结局的影响。方法前瞻性分析行胚胎植入前染色体非整倍体检测(PGT-A)助孕的134例患者,其中30例行自身对照,104例行组间对照。根据是否添加GH分为GH预处理组和GH非预处理组,GH预处理为促性腺激素(Gn)启动前行4~6周的GH 2 U/d皮下注射,Gn启动日剂量加倍直至扳机日,GH非预处理为未用过GH处理。前次PGT-A周期失败后一年内再次行PGT-A时予GH预处理构成自身对照组。组间对照和自身对照分别比较各组间的基本情况、囊胚情况及助孕结局。结果无论组间对照还是自身对照,GH预处理后的HCG日子宫内膜厚度、卵巢敏感指数(OSI)、获卵数、MII卵数、2PN数、2PN受精率、可利用卵母细胞率、活检囊胚数、整倍体囊胚数、整倍体囊胚率、至少一个整倍体率均明显增加,差异均有统计学意义(P<0.05),GH预处理后的Gn总量、Gn天数、嵌合体囊胚数、嵌合体囊胚率无明显变化,差异均无统计学意义,GH预处理后的种植率、临床妊娠率均有提高,但差异均无统计学意义。结论GH预处理可以明显增加行PGT-A患者的整倍体数和整倍体率,并有改善妊娠结局的趋势。

女性体质量指数与胚胎倍性及临床结局的相关性

目的:分析女性的体质量指数(BMI)是否影响胚胎倍性,并探讨其与整倍体胚胎移植临床结局的相关性。方法:共纳入随机分配至胚胎植入前非整倍体遗传学检测(PGT-A)组的606例不孕症女性,根据BMI将PGT-A患者分为4组:体重过低(<18.5 kg/m^(2),45例)、正常(18.5~24.9kg/m^(2),407例)、超重(25~29.9kg/m^(2),130例)和肥胖(≥30kg/m^(2),24例)。分析四组患者基于二代测序(NGS)技术的PGT-A检测结果、整倍体胚胎移植妊娠结局、新生儿结局及妊娠期并发症。结果:BMI<18.5kg/m^(2)、18.5~24.9 kg/m^(2)、25~29.9kg/m^(2)和≥30kg/m^(2)组的整倍体率分别为70.37%、69.63%、70.00%和69.44%,累积活产率分别为82.22%、77.39%、75.38%和75.00%,差异均无统计学意义(P>0.05)。调整潜在混杂因素后,各组间的胚胎倍性及临床结局各项指标比较,差异无统计学意义(P>0.05)。结论:在预后良好人群中,女性BMI与胚胎倍性和临床结局无明显相关性。

小鼠囊胚培养液用于染色体筛查的研究

目的:利用小鼠囊胚培养液探讨无创胚胎植入前非整倍体筛查(niPGT-A)的可行性,为优化和发展无创胚胎植入前遗传学筛查技术提供理论依据和数据支持。方法:本研究共收集11枚小鼠囊胚,同一囊胚分别取少量滋养外胚层细胞(TE)作为金标准、内细胞团(ICM)细胞作为阳性对照、胚胎培养液(BCM)作为待验证样本。所有胚胎样本基于全基因组测序数据进行染色体状态分析,并比较同一胚胎不同样本间染色体倍性一致性。结果:在11枚小鼠囊胚中,BCM、TE和ICM的PGT-A检出率分别为82.8%(9/11)、72.7%(8/11)和100%(11/11)。BCM与TE、BCM与ICM及TE与ICM的染色体一致性分别为85.7%(6/7),88.9%(8/9),87.5%(7/9),任意两组间差异无统计学意义(P>0.05)。结论:在小鼠模型中,相比于同期活检的滋养外胚层细胞,利用囊胚培养液中的游离DNA行无创胚胎植入前非整倍体检测可获得更高的检出率和更为准确的染色体检测结果。

Current Status and Recent Advances in Preimplantation Genetic Testing for Structural Rearrangements

Preimplantation genetic testing(PGT)is an early form of prenatal genetic diagnosis,which can identify the abnormal embryos cultured in vitro,allow only transfer of genetically normal embryos,and improve the pregnancy rate.In recent years,the rapid development of microarrays and next-generation sequencing(NGS)technologies has remarkably accelerated the clinical application of PGT.In particular,a variety of detection methods have emerged and achieved significant progress in PGT for structural rearrangements(PGT-SR).The detection-related abilities of these methods range from the detection of limited chromosome aneuploidy to comprehensive chromosome screening of the whole genome to differentiation of embryos with normal or balanced translocation/inversion karyotypes.In this study,we reviewed PGT-SR-related detection techniques to provide a better reference for clinical application and research.We have also discussed the potential development of novel techniques in the future.

内膜准备方案对PGT-A患者首次整倍体单囊胚移植妊娠结局及产科结局的影响

目的探讨不同内膜准备方案对胚胎植入前非整倍体检测(preimplantation genetic testing for aneuploidies,PGT-A)后整倍体单囊胚移植妊娠结局及产科结局的影响。方法回顾性分析2015年9月至2021年7月期间于郑州大学第一附属医院生殖医学中心行PGT-A助孕后整倍体单囊胚移植患者的临床资料,根据内膜准备方案分为自然周期组(n=80)和激素替代周期组(n=259),比较两组患者妊娠结局及产科结局的差异,并通过二元logistic回归探究影响PGT-A患者妊娠结局及产科结局的影响因素,再将年龄分层进一步探究不同内膜准备方案的妊娠结局。结果与自然周期组相比,激素替代周期组优质囊胚率显著升高,但活产率降低、流产率升高(P<0.05),两组临床妊娠率比较差异无统计学意义(P>0.05)。激素替代周期剖宫产率和巨大儿发生率高于自然周期组,低体重儿发生率低于自然周期组,但差异无统计学意义(P>0.05)。二元logistic回归显示:女方年龄是流产的独立影响因素,女方年龄和囊胚发育天数是活产的独立影响因素(P<0.05),内膜准备方案并不是妊娠及产科结局的影响因素(P>0.05)。进一步分析结果显示:35岁以上患者,自然周期组的活产率显著高于激素替代周期组,而流产率显著低于激素替代周期组(P<0.05)。结论内膜准备方案并不影响PGT-A助孕后整倍体单囊胚移植患者的妊娠结局及产科结局。但在高龄患者中,采用自然周期方案可获得更高的活产率、更低的流产率。

胚胎植入前染色体非整倍体检测在不同适应证中的助孕结局分析

目的 分析胚胎植入前染色体非整倍体检测(PGT-A)在不同适应证中的助孕结局。方法 回顾性分析PGT-A助孕的549对夫妻,按PGT-A适应证分6组,反复妊娠丢失(304例)、反复种植失败(57例)、高龄(38岁及以上,80例)、不良妊娠史(绒毛染色体三体或不良妊娠,24例)、男性因素不育(67例)、性染色体数目异常(17例),比较基本情况、获卵及胚胎活检情况、妊娠结局。结果 各组之间平均年龄、促性腺激素(Gn)使用天数差异有统计学意义(P<0.001)。各组之间平均获卵数、优质胚胎率、嵌合胚胎率、异常胚胎率、胚胎活检正常率及平均卵巢敏感性指数(OSI)差异有统计学意义(P=0.03、P<0.001、P=0.03、P<0.001、P<0.001、P<0.001);高龄组异常胚胎率在6组中最高,平均获卵数、平均OSI及胚胎活检正常率最低。6组每取卵周期临床妊娠率及持续妊娠率、累积妊娠率差异有统计学意义(P<0.001),6组每移植周期临床妊娠率及持续妊娠率、除男性因素不育组外其余5组之间累积妊娠率差异无统计学意义。结论 PGT-A可检出整倍体胚胎移植,提高妊娠效率;高龄人群有正常胚胎移植,也可获得较好的妊娠率,可能缩短其“抱婴回家”时间。同时,PGT-A可大幅提高因男性因素不育人群的妊娠结局。

D3胚胎卵裂球数目与胚胎发育潜能和整倍性的相关性研究

目的通过高通量测序技术(NGS)探讨D3胚胎卵裂球数目与胚胎发育潜能和整倍性之间的关系。方法回顾性分析2018年1月至2021年12月于本院生殖中心行胚胎植入前非整倍体遗传学检测(PGT-A)助孕的261个治疗周期共920枚活检囊胚的数据资料,根据D3胚胎卵裂球数目不同进行分组(4细胞组、5细胞组、6细胞组、7细胞组、8细胞组、9细胞组、10细胞组及≥11细胞组),评估各组后续获得优质囊胚和整倍体囊胚的机会,并统计分析各组间临床结局的差异。结果(1)与8细胞组相比,5细胞、6细胞、7细胞及10细胞组优质囊胚率显著降低(P=0.00),调整女方年龄因素以后,差异仍有显著性(P=0.00);卵裂球数目合并分组后,≤7细胞组、9~10细胞组的优质囊胚率显著低于8细胞组,调整年龄因素以后,差异仍有显著性(P<0.05);≥11细胞组优质囊胚率与8细胞组间无显著性差异(P=0.51),调整女方年龄后,仍无显著性差异(P=0.45)。(2)与8细胞组相比,其他各组囊胚的整倍体率均无显著性差异(P>0.05);调整年龄后,各组间囊胚整倍体率仍无显著性差异(P>0.05);卵裂球数目合并分组后,各组间随着卵裂球发育速度增快,囊胚整倍体率增加,但差异尚无统计学意义(P>0.05);调整年龄混杂因素后,各组间的囊胚整倍体率仍无显著性差异(P>0.05)。(3)随着卵裂球数目增加,各组临床妊娠率、活产率呈上升趋势;卵裂球数目合并分组后,≥11细胞组的临床妊娠率和活产率最高,≤7细胞组的临床妊娠率和活产率最低,但差异均尚无统计学意义(P>0.05)。结论D3卵裂期胚胎的发育潜能随着卵裂球数目的增加而增加,≥11细胞的D3快速分裂胚胎具有同8细胞胚胎相似的整倍体率和活产率,为临床单胚胎移植胚胎选择策略提供一定的参考。

嵌合体胚胎移植的临床研究现状

嵌合体胚胎是介于整倍体胚胎和非整倍体胚胎之间的第3种胚胎,是包含2种及2种以上遗传学不同细胞系的胚胎。随着遗传学技术的发展,二代测序技术对嵌合体胚胎的检出准确率更高。嵌合体胚胎移植结局的不确定性较高,着床失败率、流产率明显高于整倍体胚胎移植,对于能否移植嵌合体胚胎目前仍存在较大争议。但嵌合体胚胎移植为无整倍体胚胎的患者提供了生育可能性,因此,大样本和长期随访研究是未来研究的热点。尽管2017—2018年,中华医学会生殖医学分会发布了植入前遗传学诊断(preimplantation genetic diagnosis,PGD)/植入前遗传学筛查(preimplantation genetic screening,PGS)指南,但仍缺乏嵌合体胚胎移植的指导意见。综述目前嵌合体胚胎移植的研究现状,旨在为临床及相关科研提供借鉴。

Outcome of Couples with Reciprocal Translocation Carrier Undergoing the First Preimplantation Genetic Testing Cycles

Background: Reciprocal translocation(RCP) causes male infertility and female recurrent pregnancy loss. Male and female carriers have different responses to meiotic disturbances. Gender difference in outcomes of the RCP couples undergoing preimplantation genetic testing(PGT) is unknown.Methods: We conducted a retrospective analysis of 238 RCP couples(124 female and 114 male carriers) divided by gender of carrier from March 2014 to March 2017. Blastocysts were divided by day 5 and day 6. Females were divided into older(≥38 years) and younger(<38 years). Logistic regression was fitted for the relationship between gender of carriers and euploidy. Euploidy rate of each group, pregnancy rate, and live birth rate between different genders were analyzed.Results: The sperm live rate, forward motile sperm rate, and normal morphology rate of serum in male RCP group were significantly decreased. The euploidy rate was 30.30% in female group and 34.90% in male group(P = 0.131); 34.50% in day 5 group and 27.50% in day 6 group(P = 0.039); 33.40% in age <38 years group and 22.40% in age ≥38 years group(P = 0.063). Day 5(odds ratio [OR] = 1.388, 95% confidence interval [CI ] = 1.012–1.904; P = 0.042) and younger age(OR = 1.753, 95% CI = 0.97–3.17; P = 0.063) were associated with euploidy. The clinical pregnancy rate(37.90% vs. 41.20%), ongoing pregnancy rate(33.10% vs. 37.70%), and live birth rate(25.80% vs. 31.60%) per initiated were not significantly different in two gender groups.Conclusions: Although gender influence is not significant, couples with male carrier showed better clinical outcomes. The embryo growing rate and female age are important predictions estimating euploidy in RCP couples.

2 654例无创产前基因检测结果分析

目的探讨无创产前基因检测在胎儿染色体非整倍体疾病诊断中的临床应用价值。方法选择在该院行胎儿染色体非整倍体无创基因检测的单胎孕妇2 654例,对孕妇外周血中游离DNA进行高通量测序,对检测结果高风险者进行羊膜腔穿刺及胎儿染色体核型分析,对检测结果阴性者进行电话随访。结果 2 654例孕妇无创基因检测结果高风险29例,包括21-三体14例,18-三体6例,47,XXY 5例,45,XO 2例,常染色体异常1例,母体染色体异常1例。对29例高风险孕妇行羊膜腔穿刺羊水细胞染色体核型分析,结果显示21-三体11例,18-三体5例,性染色体异常4例。结论无创产前基因检测在诊断胎儿染色体非整倍体异常有较高的特异性和准确性,有较高的临床应用价值,但存在一定的假阳性,应掌握指征。

9号染色体倒位携带对于胚胎染色体及妊娠结局的影响

目的分析因复发性流产(RSA)行胚胎植入前筛查(Preimplantation genetic test for aneuploidies,PGT-A)包括9号染色体倒位[inv(9)]携带者及非inv(9)携带者胚胎检测结果及妊娠结局,探讨inv(9)携带对于胚胎染色体及妊娠的影响。方法回顾性分析2007年9月至2018年6月我中心就诊的因RSA行PGT-A的165对夫妇。夫妇双方至少有一位染色体核型为inv(9)的30对夫妇作为A组,染色体核型正常的135对夫妇作为B组;比较A组、B组胚胎检测结果以及妊娠结局。结果一次取卵周期中,A组总移植周期为34周期,每取卵周期累积妊娠率为76.9%,流产率为25.0%;B组总移植周期为125周期,每取卵累积妊娠率为74.0%,流产率为16.9%;两组间妊娠率及流产率均无统计学差异(P>0.05),但A组胚胎种植率显著低于B组(41.7%vs.61.4%)(P<0.05)。A组检测胚胎数209枚,胚胎染色体异常率为62.2%,但异常染色体均非9号染色体;B组活检胚胎数532枚,胚胎染色体异常率51.5%,显著低于A组(P<0.05)。结论 inv(9)有增加其它染色体异常的趋势且可能影响胚胎种植,导致RSA风险增加。

优化样品留取方法可提高囊胚培养液中游离DNA整倍性的检测效率

目的探讨利用囊胚培养液中游离DNA检测胚胎整倍性的样品留取优化方案。方法本研究共纳入239枚囊胚,所有的囊胚均来源于北京大学第三医院生殖中心胚胎植入前遗传学整倍体检测技术(PGT-A)周期。纳入患者的PGT-A指征为女方高龄(>38岁)、复发性流产、反复着床失败。胚胎在D3天更换囊胚培养液,并采取单胚胎单独培养,收集可以PGT-A活检囊胚的培养液。收集方法根据是否胚胎打孔以及胚胎培养时长分为三个阶段。PGT-A囊胚活检的滋养层细胞采用SNP芯片检测整倍性;培养液游离DNA采用二代测序方法(NGS)进行无创胚胎染色体整倍性筛查(NiPGT-A);背靠背比较两者的结果,分析NiPGT-A方法的检出率、NiPGT-A结果与PGT-A结果倍性一致性、整倍体符合率、非整倍体符合率、颗粒细胞污染率等。共有56枚PGT-A整倍性胚胎移植,分析对应胚胎的NiPGT-A结果,统计临床妊娠结局。结果239枚囊胚中209枚囊胚有NiPGT-A结果,检出率为87.5%。随着采样方法的改进,NiPGT-A结果与PGT-A结果倍性一致性、整倍体符合率逐渐提高;颗粒细胞污染率逐渐降低。239枚胚胎中有56枚PGT-A整倍体的胚胎移植,23枚胚胎的结局为活产/持续妊娠。NiPGT-A没有结果的胚胎妊娠率较高(71.4%),高于NiPGT-A为整倍体或者非整倍体胚胎的临床妊娠结局(33.3%,40.9%)。结论囊胚培养液的样品留取方法对于NiPGT-A检测至关重要,推荐D3天再次去除颗粒细胞、采集D4-D5/6天囊胚培养液,而且不要针对胚胎进行打孔处理。

无创胚胎植入前非整倍体筛查的诊断有效性分析

【目的】探讨无创胚胎植入前非整倍体筛查(niPGT-A)的诊断有效性。【方法】收集并再次分析了2018年7月到2019年7月期间,在中山大学附属第六医院生殖医学中心因染色体相互易位,或罗氏易位经首次滋养层细胞(TE)活检(TE1)行植入前,遗传学检测被诊断为非整倍体或嵌合的患者捐献胚胎24枚。解冻捐赠胚胎行滋养层细胞活检(TE2)、内细胞团活检(ICM)并收集囊胚培养液/囊胚腔液(BCM/BF)通过高通量测序技术获得遗传信息。以ICM的结果为胚胎真实染色体结果,比较TE1、TE2及BCM/BF的核型一致性。【结果】TE1、TE2及BCM/BF与相应的ICM的核型一致性分别为66.7%,87.5%和79.2%(P>0.05),任意两组之间的差异没有统计学意义(P>0.05)。【结论】利用囊胚培养液/囊胚腔液中的游离DNA行无创胚胎植入前非整倍体筛查可以获得与同期活检的TE2和ICM相似的诊断有效性。

囊胚培养液中游离DNA的重要染色体整倍性研究

目的探讨利用培养液中游离DNA检测对应囊胚的13、15、16、18、21、22号染色体整倍性的效率。方法本研究共纳入239枚囊胚,所有的胚胎均来源于PGT-A周期。囊胚活检的滋养层细胞采用SNP芯片检测胚胎的整倍性;培养液游离DNA采用无创胚胎染色体筛查(NICS)方法检测整倍性;采用盲法比较两者的结果。计算NICS在检测13、15、16、18、21、22号染色体整倍性的敏感性、特异性、诊断效率、阳性预测值和阴性预测值。结果239枚囊胚中的209枚有NICS结果,NICS检出率为87.5%(209/239)。NICS方法在检测13、15、16、18、21、22号染色体的特异性、效率比较高,均>90%,NICS结果的阴性预测值超过97%;但是敏感性波动较大(33%~100%),阳性预测值较低(<50%)。结论NICS方法可以检测囊胚整倍性。当NICS结果显示这些重要染色体(13、15、16、18、21、22号染色体)为整倍体时,对应囊胚该条染色体整倍性几率高;但是当NICS结果显示这些重要染色体为非整倍体时,对应胚胎并不一定为非整倍体。

下一代测序技术在遗传病临床检测中的应用

DNA测序技术已广泛应用于生物学研究,很多生物学问题也都可以借助测序技术予以解决。过去10年(2005-2015年),下一代测序技术(next generation sequencing,NGS)逐步从新技术成长为主流的测序技术,并逐渐走进临床诊断领域。NGS以其简单、快速、高分辨率、高通量的特点,在传染性疾病防控、肿瘤的早诊早治、遗传病的早期筛查和诊断、无创产前筛查、胚胎植入前诊断和筛查等领域发挥越来愈大的作用,已成为目前临床领域最具有应用前景的技术之一。同时下一代测序技术领域仍在快速发展,新的测序技术及数据分析技术还在不断涌现,序列数据库和测序数据快速增加。下一代测序技术也有助于以更低廉的价格,更全面、更深入地来分析基因组、转录组及蛋白质相互作用组的各项数据。可以预见,各种测序将成为一项广泛使用的常规实验手段,有望给生物医学研究领域和医学临床诊断带来革命性的变革。本文主要针对下一代测序技术在无创产前诊断、遗传病检测、胚胎植入前诊断和筛查中的临床应用进行介绍和评述。