高通量测序法胚胎植入前染色体结构变异检测评价  

Evaluation of preimplantation genetic testing for structural rearrangements based on next generation sequencing

作　　者：孙楠[1] 许建霞[1] 赵丁丁 张文新 孔令印 黄杰[1] 曲守方[1] SUN Nan;XU Jianxia;ZHAO Dingding;ZHANG Wenxin;KONG Lingyin;HUANG Jie;QU Shoufang(National Institutes for Food and Drug Control,Beijing,China,100050;Basecare Medical Device Co.,Ltd.,Suzhou,Jiangsu,China,215000)

机构地区：[1]中国食品药品检定研究院,北京100050 [2]苏州贝康医疗器械有限公司,江苏苏州215000

出　　处：《分子诊断与治疗杂志》2024年第6期993-996,共4页Journal of Molecular Diagnostics and Therapy

基　　金：国家重点研发计划项目(2021YFC2700500)。

摘　　要：目的 使用建立的染色体结构异常的家系样本,评价基于高通量测序法的胚胎植入前染色体结构异常检测试剂盒的性能。方法 首先将家系中模拟胚胎细胞样本进行单细胞全基因组扩增,然后扩增产物和家系中其他样本的DNA进行酶切打断、接头连接和磁珠富集等操作完成文库构建。将文库进行定量,使用高通量测序仪进行测序。使用生物信息学软件把测序得到的序列与人基因组参考序列比对,并进行SNP分析。结合家系亲缘关系筛选出有效SNP位点,构建家系单体型图谱。最后对易位断点上下游紧密连锁的区域进行分析,鉴别出易位携带型胚胎和正常型胚胎。结果 参考样本的测序染色体拷贝数变异(CNV)结果,确定1号染色体断点位置为chr1:194390001,胚胎样本的分型有效区域为断点上游1 Mb区域(chr1:193390001-194390001),有63个有效位点;11号染色体断点位置为chr11:133780001,胚胎样本分型有效区域为断点上游1Mb区域(chr11:132780001-133780001),有36个有效位点。通过单体型分型判断胚胎样本是携带型。结论 评价的胚胎植入前染色体结构异常检测试剂盒能够准确检出染色体结构异常的家系样本,为临床提供了一种新的染色体结构变异检测方法。Objective To evaluate the performance of preimplantation genetic testing(PGT)kit for structural rearrangements based on next generation sequencing using pedigrees.Methods First,we con-ducted single-cell whole-genome amplification of simulated embryonic cell samples.Next,we constructed the li-brary by performing enzyme digestion,splicing,and magnetic bead enrichment with DNA from samples in the pedigrees.We quantified the library and used a sequencer for sequencing.We then used bioinformatics software to align the acquired sequence with the human genome reference sequence and conducted SNP analysis.Based on the genetic relationship of the pedigrees,we screened effective SNP loci and constructed a haplotype map of the pedigrees.Finally,we analyzed the regions closely linked upstream and downstream of the translocation break-point to identify embryos carrying the translocation and normal embryos.Results According to the sequencing results for copy number variation(CNV)of the reference sample,the breakpoint position of chromosome 1 was chr1:194390001.The effective typing region of the embryonic sample was the upstream 1 Mb region(chr1:193390001-194390001)with 63 effective sites.The breakpoint position of chromosome 11 was chr11:13378000.The effective typing region of the embryonic sample was the upstream 1 Mb region(chr11:132780001-133780001)with 36 effective sites.The embryo sample was classified as a carrier type through haplotype clas-sification.Conclusion The evaluated preimplantation chromosomal structural abnormality detection kit can accurately detect chromosomal structural abnormalities in family samples,offering a new method of detection.

关 键 词：平衡易位 罗氏易位 胚胎植入前遗传学检测 结构变异 单体型连锁分析 

分 类 号：R714.8[医药卫生—妇产科学]