应用PCR-SSP方法快速检测基因HLA-B* 1502

Rapid Detection of HLA-B*1502 Allele Using PCR-SSP Method

目的通过序列特异性引物-聚合酶链反应(PCR-SSP)方法检测等位基因HLA-B*1502,并对检测结果进行讨论。方法根据HLA核苷酸碱基序列的多态性和已知的DNA序列,设计一系列等位基因型特异性引物,通过特定PCR反应体系扩增各等位基因型特异性DNA片段,产生相对应的特异性扩增产物条带,并用琼脂糖凝胶电泳检测PCR产物。结果运用4对等位基因型特异性引物,用PCR仪扩增出2份样本等位基因型HLA-B*1502,并与阴性和空白样本平行对照。结论应用PCR-SSP方法检测基因HLA-B*1502,快速、简便、安全、有效。

Objective To establish a PCR-SSP method for rapid detection of HLA-B ＊ 1502 allele and to analyze the results. Methods A set of 4 sequence-special primers and a pair of internal control primers were de- signed and synthesized according to the polymorphism of HLA nucleotide base sequence and the known sequence of DNA. A special system of PCR was used to amplify the restriction fragment of DNA in order to produce a stripe product of DNA. Using the electrophoresis of agarose gel, the amplifyied product of PCR-SSP was identified. Results A high-resolution PCR-SSP was established and two samples of HLA-B ＊ 1502 allele were successfully typed. Con- clusion This PCR-SSP technology can rapidly detect HLA-B ＊ 1502 allele and has the advantages of easier distin- guishablity, stronger specificity,lower cost, and better suitablity for clinical application.