本文選題：QT延長(zhǎng)綜合征 + 高通量核苷酸測(cè)序　； 參考：《中國循環(huán)雜志》2017年08期

【摘要】：目的:探討二代測(cè)序法在先天性長(zhǎng)QT綜合征(LQTS)臨床基因檢測(cè)中的假陰性問題。方法:選取2個(gè)商業(yè)醫(yī)學(xué)檢驗(yàn)實(shí)驗(yàn)室(Lab1和Lab2,Hi Seq2000測(cè)序平臺(tái))、1個(gè)商業(yè)科研服務(wù)實(shí)驗(yàn)室(Lab3,Ion Torrnet測(cè)序平臺(tái))和1個(gè)學(xué)術(shù)機(jī)構(gòu)實(shí)驗(yàn)室(Lab 4,Hi Seq2000測(cè)序平臺(tái))產(chǎn)生的共28例樣本數(shù)據(jù)(Lab1:6例;Lab2:8例;Lab3:8例;Lab4:6例),定量分析LQTS的三個(gè)主要致病基因KCNQ1、KCNH2和SCN5A外顯子區(qū)域測(cè)序覆蓋度以及可能漏檢的致病變異數(shù)目。結(jié)果:采用Hi Seq2000測(cè)序平臺(tái)的3個(gè)實(shí)驗(yàn)室(Lab1、Lab2和Lab4)中,三個(gè)致病基因外顯子區(qū)域覆蓋度10倍的比例均高于98%,覆蓋度30倍的區(qū)域介于90%~95%。KCNQ1在兩個(gè)商業(yè)醫(yī)學(xué)檢驗(yàn)實(shí)驗(yàn)室的14例樣本中,低于10倍和30倍覆蓋的外顯子區(qū)域比例平均為3.63%和9.84%;低于10倍覆蓋區(qū)域集中在第一外顯子,平均包含約2%的已知致病或疑似致病變異。KCNH2在兩個(gè)商業(yè)醫(yī)學(xué)檢驗(yàn)實(shí)驗(yàn)室14個(gè)樣本中,低于10倍和30倍覆蓋的區(qū)域分別為2.64%和15.76%,低覆蓋區(qū)分布在多個(gè)外顯子中。Lab1的數(shù)據(jù)中,KCNH2低于30倍覆蓋區(qū)域最高達(dá)28.56%,其內(nèi)包含已知致病或疑似致病變異113個(gè)(19.79%)。SCN5A的整體覆蓋度最好,四個(gè)實(shí)驗(yàn)室的數(shù)據(jù)都不存在低于10倍覆蓋的區(qū)域,其中兩個(gè)商業(yè)醫(yī)學(xué)檢驗(yàn)實(shí)驗(yàn)室也不存在低于30倍覆蓋的區(qū)域。結(jié)論:當(dāng)前的LQTS基因二代測(cè)序檢測(cè)中,KCNQ1和KCNH2都存在一定程度的低覆蓋區(qū),因此普遍存在漏檢致病變異的可能,假陰性問題值得高度重視。
[Abstract]:Objective: to investigate the false negative of second generation sequencing in clinical gene detection of congenital long QT syndrome (LQTS). Methods: a total of 28 samples were collected from two commercial medical laboratory laboratories: Lab1 and Lab2Hi Seq2000 sequencing platform, 1 commercial research service laboratory Lab3 + Ion Torrnet sequencing platform, and 1 academic laboratory lab, Lab4Hi Seq2000 sequencing platform. Six cases of Lab1: 8 cases of Lab3: 8 cases of Lab4: 6 cases of LQTS quantitative analysis of the three main pathogenic genes KCNQ1KCNH2 and SCN5A exon region sequencing coverage and the number of possible misdetected pathogenic variation. Results: in the three laboratories using the Hi Seq2000 sequencing platform, the proportion of the three pathogenic gene exon region coverage was 10 times higher than that of 98%, and the area with 30 times coverage was between 90%~95%.KCNQ1 in 14 samples from two commercial medical laboratory. The proportion of exon regions less than 10 times and 30 times of coverage averaged 3.63% and 9.84% respectively, and the areas below 10 times coverage were concentrated in the first exon, containing on average about 2% of known pathogenic or suspected pathogenic variations .KCNH2 in 14 samples from two commercial medical laboratory laboratories. The areas covered by less than 10 times and 30 times were 2.64% and 15.76%, respectively. In the data of .Lab1 of multiple exons, the coverage area of KCNH _ 2 was lower than 30 times, the highest was 28.566.The overall coverage of the area containing 113 known pathogenic or suspected pathogenic variations was the best, and the overall coverage of SCN5A was the best. No area of less than 10 times coverage exists in all four laboratories, and no area less than 30 times coverage exists in two of the commercial medical laboratories. Conclusion: in the second generation sequencing of LQTS gene, both KCNQ1 and KCNH2 have low coverage to some extent. Therefore, the possibility of missing the pathogenic variation is common, and the false negative problem should be paid more attention to.
【作者單位】： 首都醫(yī)科大學(xué)附屬北京安貞醫(yī)院心內(nèi)科國家心血管病臨床醫(yī)學(xué)研究中心;
【基金】：國家自然科學(xué)基金(81500246,81470465) 北京市自然科學(xué)基金(7161003) 北京市醫(yī)管局臨床醫(yī)學(xué)發(fā)展專項(xiàng)(ZYLX201302)
【分類號(hào)】：R541.7
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本文編號(hào)：1969098