多重聚合酶链反应对缺失型α地中海贫血的诊断

Geneticdiagnosis of α-thalassemia deletion by multiplex polymerase chain reaction (PCR)

目的 探讨一种简便、快速可同时检测东南亚缺失 ( SEA)、右侧缺失 ( α3 .7)和左侧缺失 ( α4.2 )地中海贫血的多重聚合酶链反应 (PCR)技术。方法 采用单管四重聚合酶链反应 (PCR)技术。结果 正常等位基因的扩增产物为 1.8kb ,东南亚缺失基因 ( SEA/αα)的扩增产物为 1.8kb和 1.3kb。右侧缺失 ( α3 .7/αα)扩增产物为 2 .0kb和 1.8kb。左侧缺失 ( α4.2 /αα)扩增产物为 1.8kb和 1.6kb。根据出现的不同条带还可判断杂合子和纯合子及缺失型HbH病。结论 本法简便、快速 ,可作为常规方法用于临床样品的分子筛查及产前诊断。

Objective To establish a rapid, simple multiplex polymerase chain reaction (PCR) method for detecting the Southeast Asia deletion(-- SEA),-α 3.7deletion and -α 4.2deletion of α-thalassemia.Methods Single barrel four-seat PCR technique was performed.Results A 1.8 kb fragment in normal chromosomes,and 2 fragments of 1.8 kb and 1.3 kb in the chromosomes with -- SEA/αα determinant,2 fragments of 2.0 kb and 1.8 kb in the chromosomes with -α 3.7/αα deletion,2 fragments of 1.8 kb and 1.6 kb in the chromosomes with -α 4.2/αα deletion were amplified.According to the differences of fragment,the heterozygous and homozygous were identified.Conclusions The established method is simple,rapid and is useful in routine laboratory for detection of carrier and prenatal diagnosis.