羊水细胞高通量测序与染色体核型分析在产前诊断中的应用研究  

A comparative study of high-throughput sequencing technology and in situ culture for prenatal diagnosis of amniotic fluid cells

作　　者：何冰[1] 黄莉[1] 张鹏[1] 王世凯[1] 黄悦悦[1] 莫伟英[1] 薛林涛[1] 田矛[2] 莫耀禧[1] 成俊萍[1] 梁羽 陈浩 

机构地区：[1]广西壮族自治区人民医院生殖医学与遗传中心,南宁530021 [2]广西壮族自治区人民医院产科,南宁530021 [3]华大基因研究院,深圳518083

出　　处：《中国临床新医学》2015年第12期1113-1116,共4页CHINESE JOURNAL OF NEW CLINICAL MEDICINE

基　　金：广西科学研究与技术开发计划课题(编号:桂科攻1140003B-66);广西自然科学基金资助项目(编号:2011GXNSFA018302);广西卫计委科研课题(编号:Z2014225;Z2014193)

摘　　要：目的通过高通量测序技术（即下一代测序技术，nextgenerationsequencing，NGS）检测孕妇羊水细胞DNA，与染色体核型分析进行对比，探索NGS在羊水细胞产前诊断中的应用价值。方法选取孕龄在18～24周之间的高龄妊娠、唐氏综合征生化筛查高风险和（或）彩超显示胎儿异常并同意产前诊断的孕妇101例，抽取孕妇羊水，提取羊水细胞DNA，制备测序文库，应用IonProton测序仪检测，所得的DNA序列与人类DNA参考数据库比对并作统计分析，并与同一样本染色体核型分析进行对照分析。结果101例羊水样本处理后经NGS技术检测判定2例染色体数目异常，37例染色体片段缺失／重复；羊水细胞培养检出2例染色体数目异常，2例9号染色体臂间倒位，2例多态性。结论利用高通量测序技术检测孕妇羊水中DNA诊断胎儿染色体数目异常，其特异性与染色体核型分析技术具有较高的一致性，并可检测出缺失／重复。染色体核型分析技术与高通量测序技术相结合在检测出生缺陷上具有较好的临床实际应用价值，并可进一步展开对疾病候选基因的研究。Objective To explore the application value of high-throughput sequencing technique（next gener- ation sequencing, NGS） in prenatal diagnosis by using NGS to test fetal free DNA in maternal amniotic fluid. Meth- ods The amniotic fluid samples from 101 pregnant women were collected. Their gestational age was between 18 and 24 weeks and had high risks of Down syndrome and/or fetal abnormalities shown by color Doppler ultrasound. The amniotic fluid samples were drawn from the pregnant women and their amniotic fluid DNAs were extracted for prepar- ing a sequencing librmT. By using Ion Proton, high-throughput sequencing procedure was carried out. The sequen- cing data were compared with the human reference gene-database and statistically analyzed. Simuhaneously, the am- niotic fluid was also collected for the chromosome karyotype analysis which compared with the NGS. Results By NGS, in 2 out of 101 cases showed chromosomal numerical abnormality, 37 cases of chromosome fragment deletion/ duplication. By the standard of chromosomal karyotypic analysis, 2 cases showed chromosomal numerical abnormali- ty, 2 cases showed chromosome 9 inversion, 2 cases showed chromosome polymorphism. Conclusion The NGS test- ed for chromosomal numerical abnormality in the maternal amniotic fluid has the same sensitivity and specificity as the chromosome karyotype analysis and can detect micro-deletion/duplication. The combination of chromosome karyotypeanalysis and high throughput sequencing technology has good clinical application prospects in the detection of birth defects.

关 键 词：高通量测序 羊水细胞 染色体 核型分析 

分 类 号：R714.5[医药卫生—妇产科学]