GAP-PCR技术快速对α-地贫血基因诊断方法的研究

RESEARCH OF GAP-PCR METHOD QUICK FOR MEDITERRANEAN ANEMIA GENE DIAGNOSIS

目的 :建立跨越断裂点的PCR技术 (也称裂口PCR、gap -PCR)为α -地贫血基因诊断提供良好的方法。方法 :应用 gap -PCR技术对 382例病人进行α -地贫血基因诊断。扩增产物带出现 10 5 3bpDNA片段时表示为α类珠蛋白基因型 (αα/αα) ,出现 743bpDNA片段增带时表示为缺失型HbH病 (α - / - - SEA) ,若样品为胎儿则为Barts水肿综合症 (- - SEA/ - - SEA) ,同时出现 10 5 3bp和 743bp两个DNA片段的扩增带表示为α -地贫杂合子 (- - SEA/αα)。结果 :382例人中 ,32 5例为正常基因型 (αα/αα) ,49例为α -地贫杂合子 (- - SEA/αα) ,8例为缺血失型HbH病 (α - / - - SEA)。结论 :gap -PCR技术应用于α -地中海贫血的血液学筛查具有简便、快速、准确的特点 。

Objective:In order to provide a kind of good method for Mediterranean anemia diagnosis PCR across gap point named gap-PCR was founded.Methods:382 persons were detected for α Mediterranean anemia gene diagnosis by gap-PCR.If 1053bp DNA segment appeared in the product the genopathy was normal α globin (αα/αα);if 743bp DNA segment showed in the product the genopathy was HbH disease (α-/-- SEA );If the samples were embryo the genopathy was Bart's edema syndrome(-- SEA /-- SEA );if 1053bp and 743bp DNA segment showed at the same time the genopathy was α Mediterranean anemia heterozygote(--SEA/αα).Results:325 persons have normal genopathy (αα/αα);49 persons have α Mediterranean anemial heterozygote (--SEA/αα)genopathy;8 persons have HbH disease(α-/-- SEA )genopathy.Conclusions:the gap-PCR is a kind of simple,quick,accurate diagnositic method for a Mediterranean anemia and can be a conventional method in place of Southern blotting.