CAJ | 학술논문

目的：探讨基于Ion Proton半导体测序平台的无创基因检测技术应用于产前诊断的可行性。方法选取2013年1至12月在郑州大学第三附属医院产检的孕龄12～32周的1000例单胎妊娠孕妇为研究对象，采用Ion Proton半导体测序平台的无创产前基因检测技术对其外周血游离胎儿DNA进行分析，其中72例行羊膜腔穿刺技术进行确诊。结果1000份标本中，染色体非整倍体高风险18例（1．8％），其中7例21-三体，4例18-三体，2例13-三体，4例性染色体异常及1例15-三体。72例孕妇经羊水穿刺确诊，胎儿21-三体、18-三体和13-三体的无创产前检出率及正确率均为100％，误诊率均为0；性染色体异常的检出率为2／2，假阳性率为1／3，1例15-三体高风险孕妇经核型确诊为假阳性。经55例核型分析和493例随访研究，该方法检测21-三体、18-三体和13-三体的特异度均为100％。采用Ion PITM芯片每次可检测12～15份标本，上机测序时间为1．5 h，整个无创检测流程为1～1．5 d。结论利用基于Ion Proton高通量测序平台的无创产前基因检测技术可快速、准确地检测出胎儿染色体非整倍体异常，该平台检测速度快、无须积累大样本量进行上机，为将来临床规范化独立开展无创性基因检测筛查胎儿染色体非整倍体异常疾病奠定了基础。
목적：탐토기우Ion Proton반도체측서평태적무창기인검측기술응용우산전진단적가행성。방법선취2013년1지12월재정주대학제삼부속의원산검적잉령12～32주적1000례단태임신잉부위연구대상，채용Ion Proton반도체측서평태적무창산전기인검측기술대기외주혈유리태인DNA진행분석，기중72례행양막강천자기술진행학진。결과1000빈표본중，염색체비정배체고풍험18례（1．8％），기중7례21-삼체，4례18-삼체，2례13-삼체，4례성염색체이상급1례15-삼체。72례잉부경양수천자학진，태인21-삼체、18-삼체화13-삼체적무창산전검출솔급정학솔균위100％，오진솔균위0；성염색체이상적검출솔위2／2，가양성솔위1／3，1례15-삼체고풍험잉부경핵형학진위가양성。경55례핵형분석화493례수방연구，해방법검측21-삼체、18-삼체화13-삼체적특이도균위100％。채용Ion PITM심편매차가검측12～15빈표본，상궤측서시간위1．5 h，정개무창검측류정위1～1．5 d。결론이용기우Ion Proton고통량측서평태적무창산전기인검측기술가쾌속、준학지검측출태인염색체비정배체이상，해평태검측속도쾌、무수적루대양본량진행상궤，위장래림상규범화독립개전무창성기인검측사사태인염색체비정배체이상질병전정료기출。
Objective To evaluate the feasibility of apply Ion Proton semiconductor sequencing platform in non-invasive prenatal genetic diagnosis .Methods Totally 1 000 pregnant women with a singleton pregnancy of 12-32 weeks gestation were selected from the Third affiliated Hospital of Zhengzhou University from Jan to Dec 2013.Using noninvasive prenatal genetic diagnosis based on Ion Proton semiconductor sequencing platform to study their cffDNA .In parallel, 72 pregnant women received invasive prenatal diagnosis by traditional chromosomal analysis with amniocentesis chorionic villus sampling .Results It′s shown that 18 out of 1 000 (1.8%) pregnant women underwent the noninvasive prenatal genetic testing had a high risk for aneuploid chromosomes , including 7 cases of 21-trisomy, 4 cases of 18-trisomy, 2 cases of 13-trisomy, 4 cases of sex chromosomal abnormality , and 1 case of 15-trisomy.It demonstrated that the rate and accuracy of fetal 21-trisomy, 13-trisomy and 18-trisomy by non-invasive prenatal genetic testing were both 100%without misdiagnosis , the rate of detection for sex chromosomal abnormality was 2/2 with a false positive rate of 1/3.However, the 15-trisomy predicted by the non-invasive prenatal diagnosis in a woman was finally proved to be a false positive .Based on the results by karyotyping (55/55) as well as follow-ups (493/493), the specificity of the non-invasive prenatal diagnosis for detection of 21-trisomy, 18-trisomy and 13-trisomy was 100%.One Ion PITM chip could detect 12 to 15 samples in 1.5 h and the whole process of noninvasive detection could be completed in 1 to 1.5 days.Conclusions The non-invasive prenatal diagnosis by Ion Proton semiconductor sequencing platform could provide fast and accurate detection of fetal aneuploidy .The benchtop high-throughput sequencing platform has laid the foundation for the independent application in clinical settings for fetal aneuploidy detection .