多重引物延伸的方法检测β-地中海贫血

Detection of β-thalassemia by the approach of multiplex primerextension

目的通过多重引物延伸的方法快速、简便地检测β-地中海贫血。方法本研究首先通过定点突变技术构建β-地中海贫血常见的5种突变位点的基因突变克隆:CD28、CD17、CD41、CD71I、VS654,然后以质粒为模板进行了引物序列、浓度、引入人工错配等条件的摸索和优化,最后采用所建立的多重引物延伸反应检测体系对临床病例样本进行检测和验证。结果本研究建立了稳定的多重引物延伸检测体系,可对以上常见的5个位点同时检测,通过对临床病例样本的验证,完全符合患者基因组测序结果。结论通过多重引物延伸检测体系的研究和摸索,作者建立了操作容易、简便、材料普通、检测花费较低的β-地中海贫血检测方法。

Objective To detect β-thalassemia by the approach of multiplex primer-extension action. Methods We constructed five kinds of clones of β-thalassemia mutation gene： CD28, CDI 7, CD4I, CD71, IVS654 by site-directed mutagenesis technique and then studied the detection conditions by the approach of multiplex primer-extension action such as the sequence and concentration of primers and artificial mismatch primers. Validated the detection system by the patient＇s sample. Result The steady system to detect β-thalassemia was established. The results of detection were according with sequencing of the patients＇ samples. Conclusion Our methods is easy, convenient, common material used, less cost to operate. So it may be useful for the convenient detection of β- thalassemia and reducing the costs of gene detection.