应用大规模平行基因组测序技术无创产前诊断胎儿常见染色体非整倍体异常  

Prenatal diagnosis of fetal chromosome aneuploidy by massively parallel genomic sequencing

作　　者：苗正友[1] 葛加美[1] 张卫华[1] 李素萍[1] 刘晓丹[1] 

机构地区：[1]浙江省嘉兴市妇幼保健院产前诊断中心,314000

出　　处：《中华医学杂志》2014年第23期1788-1790,共3页National Medical Journal of China

基　　金：浙江省科技计划（2013C33108）

摘　　要：目的：探讨大规模平行基因组测序技术检测孕妇外周血浆中游离DNA，行胎儿染色体非整倍体的无创产前诊断，在检测胎儿染色体拷贝数的可行性。方法选择2012年2月至2013年7月在浙江省嘉兴市妇幼保健院产前诊断中心就诊，孕龄在13～33周，唐氏血清学筛查高风险、高龄孕妇或超声显示有异常的孕妇1264例，行无创产前诊断。抽取孕妇外周静脉血，提取血浆DNA，制备测序文库，采用Illumina HiSeq2000测序平台对孕妇外周血中游离胎儿DNA进行测序分析，对检测结果阳性者行介入性穿刺及胎儿染色体分析；对检测结果阴性者随访。结果1264例孕妇中，检测结果阳性者共20例。与介入性诊断染色体核型分析的结果相对照，13例21-三体检测阳性者，诊断符合率100％。6例18-三体检测阳性者中，有5例进行了羊膜腔穿刺（1例发生死胎除外），其中4例为18-三体，1例为正常核型，经检测为胎盘18-三体嵌合。13-三体1例，与核型分析相符。结论大规模平行基因组测序技术行无创产前诊断胎儿染色体非整倍体，可以作为血清学筛查高风险孕妇及其他有产前诊断指征孕妇的进一步筛选方法，可大大减少介入性产前诊断的数量。其敏感性、特异性与染色体核型分析技术具有较高的一致性。Objective To perform non-invasive prenatal diagnosis （ NIPD ） of chromosome aneuploidy by detecting free DNA in maternal peripheral plasma by massively parallel genomic sequencing and determine the feasibility of detecting cell-free fetal DNA in chromosomal copy.Methods The plasma samples from 1 264 gravidas were collected from February 2012 to July 2013 at our center.Those pregnant women with a gestational age of 13 to 33 weeks having high risks in serological screening or aged over 35 years or abnormal fetus on ultrasonography.The peripheral venous blood samples were drawn from pregnant women and plasma DNA was extracted for preparing a sequencing library.By using Illumina HiSeq2000, high-throughput sequencing was performed.Amnocentesis and karyotypic analysis were conducted for positive cases and those with negative results were followed up.Results By massively parallel genomic sequencing , 20 of them showed positive results.By the standard of chromosomal karyotypic analysis , there were 21 positive cases of 13 trisomy and the diagnostic accordance rate was 100%.Among 18 positive cases of 6 trisomy, there were amniocentesis （n=5, except for one dead fetus）, trisomy 18 （n=4） and normal karyotype with placenta chimera （n=1）.One positive case of 13 trisomy was in accordance with karyotypic analysis.Conclusion Massively parallel sequencing for NIPD may be used as further screening for pregnant women with high risks of serological screening.And this technology is highly consistent with karyotypic analysis in terms of sensitivity and specificity.

关 键 词：唐氏综合征 产前诊断 DNA 高通量测序 非整倍体 

分 类 号：R714.55[医药卫生—妇产科学]