两步多重聚合酶链反应对假肥大型肌营养障碍的基因诊断

作者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(mPCR扩增dystrophia基因的9对DNA序列。对19例Duchenne型肌营养不良(DMD)和2例Becker型肌营养不良(BMD)进行基国诊断。首先用缺失率较高的5对引物扩增,检出缺失者8例,再用4对引物扩增,检出缺夫者2例。这样,9 对引物多重PCR总缺失率占受检患者的47.2％,表明可检测出90％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和(或)BMD的基因诊断。此法简便,快速又免除了使用同位素的困扰,不失为一种对DMD和(或)BMD快速诊断的好方法。

Two-step multiplex polymerase chain reaction for gene diagnosis of progressive pseudohypertrophic muscular dystrophy

In the present study. 9 oligonucleotide primers were employed to amplify 9 pairs of DNA sequences of dystrophia gene using two-step multiplex polymerase chain reaction (mPCR). Furthermore, gene diagnosis was undertaken of 19 cases of Duchenne muscular dystrophy (DMD) and 2 cases of Becker muscular dystrophy. The findings showed that 8 cases of deletion were detected by 5 amplified 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 mPCR was 47. 2% of the patients examined, suggesting that about 90% of patients with gene deletion were delected. The results of the experiment show that two step mPCR can be used in the gene diagnosis of DMD/BMD. This method is not only simple, convenient and rapid, but also free radioisotope interforence. Therefore, this method may be of importance for rapid diagnosis of DMD/BMD.