用于胚胎植入前遗传学检测安全性保障的分子标签体系构建  

Construction of a DNA molecular tag system for the safety assurance of genetic testing of preimplantation embryos

作　　者：黄志清 丁韬力 林昕迪 史航 余爱丽[1] 宋丽 孙艳 郑备红[1] HUANG Zhi-qing;DING Tao-li;LIN Xin-di;SHI Hang;YU Ai-li;SONG Li;SUN Yan;ZHENG Bei-hong(Center of Reproductive Medicine,Fujian Maternity&Child Health Hospital,Fuzhou 350001;Yikon Genetics Inc.,Suzhou 215127;Fujian Maternal-Fetal Clinical Medicine Research Center,Fuzhou 350001)

机构地区：[1]福建省妇幼保健院生殖医学中心,福州350001 [2]苏州亿康医学检验有限公司,苏州215127 [3]福建省母胎医学临床医学研究中心,福州350001

出　　处：《生殖医学杂志》2024年第12期1571-1577,共7页Journal of Reproductive Medicine

基　　金：福建省卫生健康科研人才培养项目青年科研课题(2019-2-23);福建省妇幼保健院科技创新启动基金博士优秀青年科学基金项目(YCXB 18-04);福建省自然科学基金青创项目(2020J05279);福建省科技创新平台项目(2021Y2012)。

摘　　要：目的 探讨采用DNA分子标签用于胚胎植入前遗传学检测(PGT)安全性保障的可行性。方法 选择2024年1—2月于福建省妇幼保健院生殖医学中心行IVF助孕的患者作实验材料来源,选择3 PN胚胎、卵裂期形态废胚、4 CC囊胚活检细胞和实际活检细胞基因组DNA扩增产物4种样本,各12例。在样本保存液管或扩增产物中预先添加特定的DNA分子标签,再进行扩增、CNV建库和NGS测序,分析全流程中间产物中的DNA分子标签序列以及NGS数据中的DNA分子标签序列。结果 PCR扩增产物的Sanger测序结果显示,均能从所有样本中测出各自唯一的DNA分子标签序列,并且均和所添加的DNA分子标签一致,也均和外部标签完全对应和一致。各样本扩增产物和测序文库浓度均在正常范围,测得的DNA分子标签序列约占所有NGS测序数据的1‰,且包括Q30在内的测序数据各质控指标也均在正常范围,按照常规生信分析得出的胚胎遗传学信息和之前测序分析的结果是完全一致的。结论 DNA分子标签可以精准有效关联冷冻的活检胚胎及其活检细胞的遗传学检测结果,还可以全流程追踪校对活检细胞的遗传学检测,从而有力保障PGT的安全性。Objective:To explore the feasibility of using DNA molecular tags in the safety assurance of genetic testing of preimplantation embryos(PGT).Methods:Four types of samples for 12 cases in each type,including tripronuclear(3PN)embryos,waste cleavage-stage embryos,biopsy samples from 4CC blastocyst,and genomic DNA amplification products of actual biopsy cells,were selected form the patients who underwent IVF assisted pregnancy at the Reproductive Medicine Center of Fujian Maternal and Child Health Hospital between January and February 2024.Specific DNA molecular tags were preadded to sample collection tubes or the amplification products before amplification,copy number variation(CNV)library construction and next-generation sequencing(NGS).The DNA molecular tag sequences in the intermediate products of the whole process and the DNA molecular tag sequences in the NGS data were analyzed.Results:The Sanger sequencing results of PCR amplification products showed that the respective unique DNA molecular tag sequences were identified in the various products of all samples,and the sequences were all consistent with the added DNA molecular tag,fully consistent with the external tag.The concentrations of the amplification products and sequencing library of each sample were within the normal range.The measured DNA molecular tag sequences accounted for only approximately 1%of all NGS sequencing data,and quality control indicators of the sequencing data,including Q30,were also within their normal ranges.The embryonic genetic information obtained through conventional bioinformatics analysis is completely consistent with the results of previous sequencing analysis.Conclusions:DNA molecular tags can accurately and effectively associate with frozen biopsy embryos and the genetic test results of biopsy cells and can also track and proofread the genetic testing of biopsy cells in the whole process,thus strongly ensuring the safety of genetic testing of preimplantation embryos.

关 键 词：辅助生殖 胚胎植入前遗传学检测 安全性 分子标签 

分 类 号：R711.6[医药卫生—妇产科学] R715.5[医药卫生—临床医学]