荧光定量聚合酶链反应联合拷贝数变异测序技术在产前诊断中的应用研究  

Application of Quantitative Fluorescent-PCR and CNV-Seq in Prenatal Diagnosis

作　　者：李琴[1] 魏兆莲[2] 乔金平[3] 陈薇[1] 方慧琴[1] 袁静[1] LI Qin;WEI Zhao-lian;QIAO Jin-ping;CHEN Wei;FANG Hui-qin;YUAN Jing(Prenatal Diagnosis Center,The First Affiliated Hospital of Anhui Medical University,Hefei 230022,China;Reproductive CenterThe First Affiliated Hospital of Anhui Medical University,Hefei 230022,China;Medical Laboratory Center,The First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)

机构地区：[1]安徽医科大学第一附属医院产前诊断中心,合肥230022 [2]安徽医科大学第一附属医院生殖中心,合肥230022 [3]安徽医科大学第一附属医院医学实验中心,合肥230022

出　　处：《国际生殖健康／计划生育杂志》2020年第6期450-455,共6页Journal of International Reproductive Health/Family Planning

基　　金：安徽省自然科学基金项目(1708085MH214)。

摘　　要：目的:回顾性分析采用荧光定量聚合酶链反应(QF-PCR)联合全基因组拷贝数变异测序(CNV-Seq)技术对270例胎儿常见染色体非整倍体进行快速产前诊断并检测>100 kb的全基因组拷贝数变异的检测结果。方法:对270例早中孕期因产前血清学筛查异常或母亲血胎儿游离DNA(无创DNA)行产前筛查结果提示高风险孕妇人群进行产前诊断的羊水样本,通过短串联重复序列(STR)位点比对后行13、18、21、X、Y染色体非整倍体的快速诊断并联合高通量DNA测序法CNV-Seq技术对染色体全基因组拷贝数变异进行检测;同时与染色体核型结果进行比较和分析。结果:QF-PCR结果中胎儿5种常见染色体(13、18、21、X、Y染色体)非整倍体共计19例,阳性率7.03%(19/270),其中21三体12例,18三体3例,XXY综合征2例,XYY综合征2例。染色体核型提示非整倍体结果与此一致。二者染色体非整倍体检出率和准确性一致。染色体核型结果中尚有11例染色体多态性。CNV-Seq异常总数43例,阳性率15.9%(43/270);其中19例为染色体非整倍体,24例微缺失微重复(明确致病CNV-Seq 9例,致病性未知CNV-Seq 5例,良性可能CNV-Seq 10例)。QF-PCR联合CNV-Seq技术检测周期平均1~2周左右,而染色体核型检测周期5周左右且有1例培养失败(后重新羊膜腔穿刺采集羊水标本)。结论:QF-PCR联合CNV-Seq技术在临床上可快速准确筛查产前高风险孕妇人群,有利于二级预防实施和减少出生缺陷,提高优生优育。Objective:To analyze retrospectively the application of quantitative fluorescent PCR(QF-PCR)combined with genome-wide copy number variation sequencing(CNV-Seq)technology in the rapid prenatal diagnosis of 270 cases of fetal common chromosome aneuploidy and the detection of genome-wide copy numbers>100 kb.Methods:A total of 270 pregnant women in the early pregnancy and the second trimester were included in this study.These women were previously showed the abnormalities of Down′s screening or the high-risk of maternal blood fetal free DNA testing(prenatal non-invasive screening).The amniotic fluid samples from these pregnant women were used for the short tandem repeated(STR)detection.After site comparison,the chromosomal aneuploidies including 13,18,21,X,Y chromosome were quickly diagnosed,and the copy number variation of the chromosomal whole-genome was detected by the combined high-throughput DNA sequencing(CNV-Seq technology).These results were then compared with the result of karyotype testing.Results:The QF-PCR test showed that there were 19 cases of aneuploid chromosomes,with a positive rate of 7.03%(19/270),which is related to five common fetal aneuploid chromosomes(13,18,21,X,Y chromosomes)including 12 cases of trisomy 21,3 cases of trisomy 18,2 cases of XXY syndrome and 2 cases of XYY syndrome.The karyotype result of aneuploidy abnormalities was suggested to be consistent with the QF-PCR test,with the same rate of aneuploidies and the same accuracy.In addition,there were 11 cases of chromosomal polymorphism in the karyotype results.There were 43 cases of CNV-Seq abnormalities,with a positive rate of 15.9%(43/270),including 19 cases of chromosomal aneuploidy abnormalities and 24 cases of microdeletions and microduplications.In these microdeletions and microduplications,there were 9 cases of pathogenic CNV-Seq,5 cases of unknown pathogenic CNV-Seq,and 10 cases of polymorphic CNV-Seq.The testing time of the QF-PCR combined with CNV-Seq technology was about 1-2 weeks,while this time of karyotype was generall

关 键 词：实时聚合酶链式反应 序列分析 DNA DNA拷贝数变异 串联重复序列 产前诊断 

分 类 号：R714.5[医药卫生—妇产科学]