基于高通量测序进行无创产前亲子鉴定的可行性

目的初步探讨基于高通量测序进行STR分型的技术方法应用于无创产前亲子鉴定的可行性。方法选择13个STR基因座(6个常染色体STR基因座,6个Y染色体STR基因座,1个性别判定基因座),进行复合PCR扩增和高通量测序文库构建后,采用Ion PGM400高通量测序平台进行测序,并采用自主研发软件NGS-STR genotyper(perl脚本)进行STR分型,本文简称上述过程为NGS-STR分型。对13个母子配对混合样本(母亲:儿子=2%~50%)、1组家系样本进行了上述NGS-STR分型,旨在(1)了解其在混合样本中的灵敏度及分型情况;(2)了解其在无创产前亲子鉴定中的应用可能性。结果 (1)当混合样本中低组分(儿子)的比例超过8%,所有基因座均可检出低组分的STR信息;(2)对1例血浆样本进行NGS-STR分型,共计69.2%的基因座可检出胎儿的STR基因型信息,且所有检出基因座均符合孟德尔遗传规律。结论初步证明了NGS-STR分型技术具有进行无创产前亲子鉴定的可行性。

Study of the feasibility of non-invasive prenatal paternity test based on next-generation sequencing

Objective To evaluate the application value of STR genotyping based on second-generation highthroughput sequencing in non-invasive prenatal paternity test.Methods 13 STR loci (six autosomal STRs,6 Y-chromosome STRs,and 1 gender determination locus) were amplifiedand the libraries were built and sequenced by using Ion PGM400 sequencing platform,and the STR genotyping wasfiledbased on the sequencing data.13 mixed samples of mother and son pairs(mother:son =2％～50％),and 1 case of maternal plasma were performed by NGS-STR genotyping in order to:1) assess its sensitivity in the mixed samples;2) assess the possibility of the application of non-invasive prenatal paternity test.Results 1) When the mixing ratio of the low sample component (son) was more than 8％,all STR information of the low component could be detected;2) for the case of plasma,NGS-STR genotypingsucceededin 69.2％ of the fetal STR loci,which proved that NGS-STR genotyping can be applied in the field of non-invasive prenatal paternity test.