摘要
应用分子生物学技术,用9组寡核苷酸引物分两步多重聚合酶链反应扩增dystrophin基因的9段脱氧核糖核酸序列。对36例DMD和4例BMD进行基因诊断。首先用缺失率较高的5对引物扩增,检出缺失者17例,再用4对引物扩增,检出缺失者2例。这样,9对引物多重PCR总缺失率占受检患者的47.5%,表明此法可检测出79.1%左右有基因缺失的患者。实验结果提示,两步多重PCR可用于DMD/BMD的基因诊断。
Abstract In the present study,9 oligonucleotide primers were employed to amplify 9 pairs of DNA sequences of dystrophin gene using the two-step multiplex polymerase chain reaction(mPCR)Furthennore,gene diagnosis were undertaken in 36 cases of Duchenne muscular dystrophy and 4 cases of Becker muscular dystrophy,The findings showed that 17 cases of deletiori were detected by 5 ampli-fied primers with a relatively high incidence of deletion and 2 more cases of deletion were detected using the remaining 4 amplified primers.The deletion rate of 9 primers using two-step mPC R was 47.5%of the patients examined,suggesting that about 79.1%of patients with gene deletion were detected.The resuIts of the experiment show that two-step mPCR can be used in the gene diagnosis of DMD/BMD.