胚胎培养液用于染色体筛查的初步研究  

Preliminary study of embryo culture medium used for chromosome screening

作　　者：韩瑞钰 王雪莹 邓佩佩 和伟 姚冠峰 郭薇 HAN Ruiyu;WANG Xueying;DENG Peipei;HE Wei;YAO Guanfeng;GUO Wei(Hebei Research Institute of Family Planning Science and Technology, Key Laboratory of the National Commission of Health and Family Planning and Healthy Birth, Shijiazhuang, Hebei Province, 050071)

机构地区：[1]河北省计划生育科学技术研究院,国家卫计委计划生育与优生重点实验室,石家庄050071

出　　处：《中国计划生育学杂志》2021年第2期308-310,共3页Chinese Journal of Family Planning

摘　　要：目的:对胚胎培养液筛查胚胎染色体进行初步探讨,为优质胚胎无创筛查体系提供数据和技术支持。方法:采集胚胎培养液86例和囊胚36例,其中56例为卵胞浆内单精子注射-胚胎移植(ICSI-ET)胚胎培养液,30例为体外受精-胚胎移植(IVF-ET)胚胎培养液。采用基因测序通用文库试剂盒对囊胚及胚胎培养液进行MALBAC单细胞全基因组扩增及文库构建,运用高通量测序平台进行测序,运用ChromGoTM2.0数据分析平台检测染色体非整倍体及10M以上的片段缺失和(或)重复。结果:86例胚胎培养液中27例未检出染色体核型,其中前期收集36例标本,检出成功率41.7%;改进方法收集50例标本,检出成功率88.0%。对36例囊胚及其培养液染色体结果进行对比,IVF-ET 18例,染色体倍性一致率38.9%,非整倍体假阳性率50.0%,假阴性率0.0%;ICSI-ET 18例,染色体倍性一致率77.8%,非整倍体的假阳性率16.7%,假阴性率5.6%。结论:ICSI-ET受精方式在胚胎培养液用于胚胎染色体的筛查中优于IVF-ET;建立完善的培养液收集流程利于胚胎染色体检测。Objective:To preliminary explore the embryo culture medium used for chromosome screening,and to provide date and technical support for the non-invasive screening system of high-quality embryos.Methods:The culture medium of 86 embryos and 36 blastocysts were collected,which included 56 embryos cultured by ICSI-ET and 30 embryos cultured by IVF-ET.Full-genome amplification and library construction of MALBAC single cells were performed on blastocysts and embryo culture media by the universal gene sequencing library kit.High-throughput sequencing platform was used for sequencing,and ChromGoTM2.0 data analysis platform was used to detect the deletion and/or duplication of chromosome aneuploidy,and fragments over 10M.Results:Chromosome karyotype was not found in culture medium of 27 of the 86 embryos,among which,36 specimens were collected in the early stage,which’s detection success rate was 41.7%.The improved method was used to collect the other 50 specimens,which’s detection success rate increased to 88%.By comparing the chromosome results of 36 blastocysts with their culture medium,a total of 18 embryos were cultured by IVF-ET,with the chromosome ploidy consistency rate of 38.9%,false positive rate of 50%and false negative rate of 0%.In 18 cases of ICSI-ET,the consistent rate of chromosome ploidy was 77.8%,the false positive rate was 16.7%,and the false negative rate was 5.6%.Conclusion:The culture medium of embryos for chromosome screening of ICSI-ET is superior to that of IVF-ET,and the establishment of culture medium collection process is benefit for embryo chromosome detection.

关 键 词：体外受精 卵胞浆内单精子注射 胚胎培养液游离DNA 无创胚胎染色体筛查技术 

分 类 号：R714.8[医药卫生—妇产科学]