泰国型α缺失地中海贫血的快速基因诊断方法的建立及其临床应用

Development and clinical application of a rapid gene diagnosis method for Thai type thalassemia

目的:建立一种快速、简便且无需核酸提取的泰国型α缺失地中海贫血的快速基因诊断方法及其在产前诊断中应用。方法:设计可以扩增α-珠蛋白基因簇中--THAI等位基因的特征序列引物组,直接以待测样本为模板,不需要核酸提取,进行聚合酶链反应(PCR)直接扩增,扩增产物进行琼脂糖凝胶电泳即可分析结果。结果:成功建立泰国缺失型地中海贫血的快速基因诊断方法,并且直接以全血、羊水、干血斑、脐带血为模板进行扩增的结果与常规跨越断裂点聚合酶链反应(gap-PCR)技术确定基因型完全一致。结论:该法不需要提取核酸,减少操作步骤与实验室污染、节约成本、快速、准确,可作为常规方法用于临床样品的地中海贫血分子基因检测及巴氏水肿胎和缺失型HbH病的产前诊断及泰国型α-地中海贫血的产前诊断新方法。

Objective:To establish a rapid,simple and non nucleic acid extraction based gene diagnosis method for Thai type Thalassemia and its application in prenatal diagnosis.Methods:The characteristic sequence Primers of-thai Allele in-globin gene cluster were designed,and the samples were directly amplified by PCR without nucleic acid extraction,the amplified products were analyzed by Agarose Gel electrophoresis.Results:A rapid genotyping method for Thai type Thalassemia was successfully established,and the results of direct amplification using whole blood,amniotic fluid,DBS and umbilical cord blood were identical with those determined by gap PCR.Conclusion:This method does not need to extract nucleic acid,reduces the operating procedure and the laboratory pollution,saves the cost,is fast,is accurate,thalassemia gene detection as a routine method for clinical samples and prenatal diagnosis of Bart’s edematous foetus and absent type HBH are expected to be new methods for clinical prenatal diagnosis of Thai type thalassemia.