基于二代测序的mini-STR分型技术应用于无创产前亲权鉴定的可行性研究  

The feasibility study of non-invasive prenatal paternity testing with mini-STRs based on next-generation sequencing

作　　者：宋文倩[1] 肖南[1] 陈玫[1] 段莹[1] 潘凌子[1] 王霓[1] 邵林楠 周世航[1] 于卫建[1] 刘铭[2] SONG Wenqian;XIAO Nan;CHEN Mei;DUAN Ying;PAN Lingzi;WANG Ni;SHAO Linnan;ZHOU Shihang;YU Weijian;LIU Ming(Dalian Blood Center,Dalian,Liaoning 116001,China;Department of Cell Biology,Dalian Medical University,Dalian,Liaoning 116044,China)

机构地区：[1]大连市血液中心,辽宁大连116001 [2]大连医科大学细胞生物学教研室,辽宁大连116044

出　　处：《中国优生与遗传杂志》2021年第6期856-862,共7页Chinese Journal of Birth Health & Heredity

基　　金：大连市青年科技之星项目(2017RQ169)。

摘　　要：目的探讨基于二代测序技术(next generation sequencing,NGS)检测孕妇血浆中胎儿短串联重复序列(short tandem repeat,STR)基因座的方法应用于无创产前亲权鉴定的可行性。方法收集28组家庭样本,首先用毛细管电泳(capillary electrophoresis,CE)法检测每组家庭STR基因型,验证父-母-婴儿的亲子关系。然后,用Illumina平台NGS测序技术检测母亲血浆中游离胎儿DNA(cell free fetal DNA,cffDNA)的23个mini-STR基因座的等位基因,筛选适合的mini-STR基因座,用于胎儿的亲权鉴定。结果对同一家庭的父亲、母亲和婴儿的STR分型结果证实28组家庭全部具有亲子关系。cffDNA样本的NGS检测结果在排除母亲DNA干扰后,与婴儿CE方法分型结果进行比对,结果显示D5S818、D19S253和D21S1270三个基因座符合率低于50%。针对余下符合率高于60%的20个基因座,将基于NGS的cffDNA的STR分型结果分别与生物学父亲和随机男性进行比对,结果显示两组一致率有显著性差异(P<0.0001),分别为75%~100%和25%~70%。结论基于NGS技术检测cffDNA的STR分型方法可应用于无创产前亲权鉴定。Objective To evaluate the feasibility of detecting cell free fetal DNA(cffDNA)from plasma of pregnant women,short tandem repeat(STR)genotyping by next generation sequencing(NGS)and applying on non-invasive prenatal paternity testing.Methods DNA samples from 28 families were collected.Paternity testing was performed by capillary electrophoresis(CE).The 23 mini-STR loci of cell free fetal DNA(cffDNA)from the maternal plasma were detected through Illumina NGS method.The loci with high accordance rate were selected and applied on prenatal paternity testing.Results It showed positive proof of paternity including father,mother and child of each family.The comparatively analysis between NGS sequencing results of cffDNA without maternal DNA interference signals and CE genotyping results of child’s DNA showed that the accordance rate of three loci including D5S818,D19S253 and D21S1270 were lower than 50%.For the other 20 loci with the accordance rate higher than 60%,the STR genotyping results showed a significant difference of matching probability between cffDNA and the DNA from the biological father(75%-100%)or from random males(25%-70%)(P<0.0001).Conclusion Our study proved that STR genotyping of cffDNA by NGS method can be applied in non-invasive prenatal paternity testing.

关 键 词：mini-STR分型 无创产前亲权鉴定 二代测序技术 血浆中游离胎儿DNA 

分 类 号：R714.5[医药卫生—妇产科学]