比较SNP-array和高通量测序在胚胎植入前遗传学检测中的效能  

Comparison of SNP-array and next generation sequencing in preimplantation genetic testing

作　　者：周红[1] 蓝月云 舒金辉[1] 汪彩珠[1] 赵鑫 梁丽芳 何升[1] 邱庆明[1] 黄朋[1] Zhou Hong;Lan Yueyun;Shu Jinhui;Wang Caizhu;Zhao Xin;Liang Lifang;He Sheng;Qiu Qingming;Huang Peng(Reproductive Center,Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region,Nanning 530013,China)

机构地区：[1]广西壮族自治区妇幼保健院生殖中心,南宁530013

出　　处：《中华生殖与避孕杂志》2023年第10期1018-1025,共8页Chinese Journal of Reproduction and Contraception

基　　金：广西医疗卫生适宜技术研究与开发项目(S201541)。

摘　　要：目的评估单核苷酸多态性微阵列(single nucleotide polymorphisms array,SNP-array)技术和第二代高通量测序(next generation sequencing,NGS)技术在胚胎植入前遗传学检测(preimplantation genetic testing,PGT)中的检测能力和效率。方法回顾性描述分析2020年1月至2022年8月期间于广西壮族自治区妇幼保健院生殖中心就诊、夫妇双方均携带有明确致病性基因突变并要求进行第三代试管婴儿助孕的188例患者资料。经卵胞质内单精子注射授精和体外培养后,共收获995枚囊胚并进行活检。患者胚胎细胞遗传物质经全基因组扩增之后,分别采用SNP-array或NGS分析其携带致病基因突变和染色体拷贝数变异(copy number variation,CNV)的情况,同时采用Sanger测序或间隙聚合酶链反应(polymerase chain reaction,PCR)对突变进行直接检测。分析女方年龄和获囊胚数的关系、胚胎携带突变和致病性CNVs的比例,比较不同分子诊断技术在PGT中的检测成功率和准确性。经遗传学检测后,选择合适的胚胎进行子宫腔内移植,并于中孕期进行羊水穿刺产前诊断。结果①成功对924个胚胎进行遗传学检测,总的检测成功率为92.9%。根据遗传学检测结果,共有389个(42.1%)胚胎可用于移植。②对胚胎进行缺失型α-地中海贫血突变检测,间隙PCR的成功率[84.9%(465/548)]低于SNP-array[98.7%(81/82)]和NGS[92.5%(431/466)]。对点突变进行检测,Sanger测序的成功率[98.5%(440/447)]与SNP-array[95.6%(110/115)]和NGS[96.1%(319/332)]的差异无统计学意义。有38个胚胎因等位基因脱扣导致直接位点检测法的结果与SNP单体型分析的结果不一致。另外,有4个胚胎因染色体重组导致SNP单体型分析失败。③与NGS相比,SNP-array检测CNV的成功率[83.7%(165/197)]较低,但是可检出更多种类的染色体拷贝数异常。④共进行了152例胚胎移植,其中有107例成功临床妊娠,有69例完成了羊水穿刺产前诊断,有42个健康儿Objective To evaluate the detection ability and efficiency of single nucleotide polymorphisms array(SNP-array)and next generation sequencing(NGS)in preimplantation genetic testing(PGT).Methods Totally 188 couples who carried pathogenic gene mutation and requested preimplantation genetic testing for monogenic(PGT-M)treatment were retrospectively analyzed in the Reproductive Center of Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region during January 2020 and August 2022.After ovulation induction,insemination was conducted by intracytoplasmic sperm injection(ICSI)and cultured in vitro,995 blastocysts were harvested and biopsied.After whole genome amplification(WGA)of the genetic material from embryonic cell samples,their carrying status of mutations and chromosome copy number variations(CNVs)were analyzed by SNP-array or NGS,respectively,and along with mutation direct detection by Sanger sequencing or Gap-PCR.The relationship between female age and the number of blastocysts was analyzed,as well as the proportion of embryos carrying mutations and pathogenic CNVs.The detection success rate and accuracy of different molecular diagnostic techniques used in PGT were compared.Amniocentesis prenatal diagnosis was performed in the second trimester after successful intrauterine transfer of embryos.Results 1)A total of 924 embryo samples were successfully performed genetic testing,with a total success rate of 92.9%,and 389 embryos(42.1%)can be transferred according to these results.2)In detecting deletionalα-thalassemia,the success rate of Gap-PCR[84.9%(465/548)]was lower than that of SNP-array[98.7%(81/82)]and NGS[92.5%(431/466)].However,the success rate of direct mutation detection by Sanger sequencing[98.5%(440/447)]was not significantly different from that by SNP-array[95.6%(110/115)]and NGS[96.1%(319/332)].There were 38 embryo samples with direct mutation detection results inconsistent with those based on SNP haplotyping.In addition,4 embryo samples failed SNP haplotyping due to chromosomal recombina

关 键 词：胚胎植入前遗传学检测 SNP单体型分析 第二代高通量测序技术 

分 类 号：R714.8[医药卫生—妇产科学]