Liu Jingzhong, Tang Ning, Liu Quanzhang, Wang Lirong, Han Han, Cai Ren, Wu Xiaoyi, Xiao Bai
Basic Medical Research Center, Beijing Chaoyang Hospital Affiliate of The Capital Medical University, Beijing, China.
Acta Haematol. 2009;122(1):17-22. doi: 10.1159/000232578. Epub 2009 Aug 15.
The prevailing cause of alpha-thalassemia in Southeast Asia is the presence of 3 deletion mutations in the alpha-globin genes (-SEA, -alpha(3.7) and -alpha(4.2)). Current detection methods include gap polymerase chain reaction (PCR), multiplex PCR and real-time PCR with SYBR Green 1 combined with dissociation curve analysis. To improve and simplify a previously published method that requires 4 separate reactions, a duplex PCR assay was designed to detect both the nondeletional and the -SEA alleles. This duplex PCR can successfully identify the nondeletional allele and both the -SEA carrier and homozygous genotypes. The combination of the duplex PCR and 2 gap PCRs (for detection of -alpha(3.7) and -alpha(4.2)) can diagnose all types of deletional alpha-thalassemia. Our method was validated by analysis of 195 DNA samples, the results of which were consistent with prior diagnoses. The developed assay can reliably diagnose alpha0-thalassemia and all types of deletional alpha-thalassemia. The diagnostic method is simple, rapid, accurate, automated, inexpensive and has a high throughput.
东南亚地区α地中海贫血的主要病因是α珠蛋白基因存在3种缺失突变（-SEA、-α(3.7)和-α(4.2)）。目前的检测方法包括缺口聚合酶链反应（PCR）、多重PCR以及结合SYBR Green 1和熔解曲线分析的实时PCR。为了改进和简化之前需要4个独立反应的方法，设计了一种双重PCR检测法来检测非缺失等位基因和-SEA等位基因。这种双重PCR能够成功鉴定非缺失等位基因以及-SEA携带者和纯合基因型。双重PCR与2种缺口PCR（用于检测-α(3.7)和-α(4.2)）相结合，可以诊断所有类型的缺失型α地中海贫血。通过对195份DNA样本的分析验证了我们的方法，其结果与之前的诊断一致。所开发的检测法能够可靠地诊断α0地中海贫血和所有类型的缺失型α地中海贫血。该诊断方法简单、快速、准确、自动化、成本低廉且通量高。
