运用单管多重聚合酶链反应检测海南黎族人群中三种缺失型α-珠蛋白生成障碍性贫血基因

目的 建立单管多重聚合酶链反应(mPCR)技术并将其运用于筛查海南黎族人群中3种最常见的缺失型α-珠蛋白生成障碍性贫血基因(α-地中海贫血基因,简称α-地贫基因)。方法 在研究成功检测3种α-地贫基因,即-^SEA、-α^3.7、-α^4.2的3个PCR反应的基础上,建立并优化单管mPCR反应的条件,检测了40例已知基因型的α-地贫患者基因,并筛查、诊断了116份海南黎族人血标本。结果 运用新建立的单管mPCR检测40例α-地贫患者DNA,所得基因型与运用3个分别的PCR技术及Southern印迹杂交技术所得到的结果一致。在116份海南黎族人中,检出-α^3.7和-α^4.2。缺失型基因的携带频率高达38．0％,且-α^4.2稍高于-α^3.7。但在该组标本中没测到-SEA。结论 单管mPCR技术准确率及灵敏度都很高,且简便易行。海南黎族人群-α^3.7和-α^4.2携带频率居国内最高。

Detection of deletional determinants of alpha-thalassemia in Li people in Hainan province by a single tube multiplex polymerase chain reaction method

OBJECTIVE: To develop a single tube multiplex polymerase chain reaction technique (mPCR) and use it to screen 3 deletional alpha-globin genes, namely -alpha(3.7), -alpha(4.2) (single gene deletion), and --SEA (South East Asia double gene deletion) in the Li people in Hainan province. METHODS: Based on previously developed 3 separate PCR methods used to detect the -alpha(3.7), -alpha(4.2), --SEA respectively, a single tube mPCR was investigated, optimized, and used to detect the genotypes of 40 samples from well-known alpha-thalassemia patients and 116 non-relative individuals of Li nationality in Hainan province. RESULTS: The results of the 40 samples detected by mPCR were the same as those by the 3 separate PCR Methods. Among the 116 sample, the combined incidence of -alpha(3.7) and -alpha(4.2) was found to be as high as 38.0%, and the -alpha(4.2) genotype was more frequent than the -alpha(3.7) genotype in the Li people. No --(SEA) deletion was found in the Li people. CONCLUSION: The mPCR technique shows a good sensitivity and specificity. The Hainan Li people has the highest incidence of -alpha(3.7) and -alpha(4.2) among Chinese.