基因拷贝数定量检测在缺失型α-地贫快速诊断中的应用  

Application of genetic copy number quantification for rapid diagnosis of alpha-thalassaemia deletion

作　　者：胥顺[1,2] 周万军[3] 牟毅 程钢[1,4] 

机构地区：[1]中山大学药学院,广东广州510006 [2]广州市达瑞抗体工程技术有限公司,广东广州510665 [3]南方医科大学基础医学院遗传教研室,广东广州510515 [4]中山大学达安基因股份有限公司,广东广州510665

出　　处：《分子诊断与治疗杂志》2014年第4期222-227,共6页Journal of Molecular Diagnostics and Therapy

基　　金：国家科技部863计划(2011AA02A109)

摘　　要：目的探讨目的基因拷贝数定量检测方法在缺失型α-地中海贫血快速诊断中的应用。方法采用四重荧光定量PCR方法检测1 385例α-地中海贫血样本的α珠蛋白基因拷贝数,同时用gap-PCR法进行平行对照检测,对结果不符样本采用MLPA法进行确认。结果 1 385例临床样本中,本方法检出αα/αα型531例、α-/αα型168例、-α/αα型93例、α-/α-型7例、-α/-α型2例、--/αα型558例、--/α-型13例、--/-α型11例、--/--型2例;其中与gap-PCR法结果不符的样本9例,MLPA复核结果显示5例检测结果一致、4例与多重荧光定量PCR结果不一致;拷贝数定量PCR法检测准确度达到99.71%,与gap-PCR检测符合率达到99.35%。结论α珠蛋白基因拷贝数定量PCR法能快速准确检测缺失型α-地中海贫血,适合大规模人群的缺失型α-地贫基因携带筛查。Objective To discuss the application of quantitative detection of HBA copy number in rapid detection of alpha-thalassaemia deletion. Methods The HBA copy number were detected by multiplex qPCR in 1 385 clinical samples, while the deletional genetypes were parallel detected by gap- PCR, and the different results with the the former two methods were confirmed by MLPA. Results In 1385 cases of clinical samples, there were 531 cases with αα/αα, 168 cases with α-/αα, 93 cases with -α/αα, 7 cases with α-/α-,2 cases with -α/-α, 558 cases with --/ctct, 13 cases with --/α-, 11 cases with --/-α, 2 cases with --/--; and 9 cases were inconsistent with the gap-PCR. The MLPA results showed that 5 cases were completely conformed by multiplex qPCR, and four cases samples did not match; the true accuracy was 99.71%, and the coincidence rate was 99.35% compared with the gap-PCR. Conclusion The HBA copy number of multiplex qPCR method, which is rapidly and accurately to detect the alphathalassaemia, is suitable for large scale population screening.

关 键 词：Α-地中海贫血 基因拷贝数 荧光定量PCR 

分 类 号：R450[医药卫生—治疗学]