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: 谭庆荣 吴保仁 王连刚
• 机 构: 西京医院神经内科,西京医院神经内科,西京医院神经内科
• 出 处: 第四军医大学学报
• 发表日期: 1993年06期
• 关键词: 基因诊断 营养障碍 肥大型 聚合酶链反应 引物扩增 基因缺失 肌营养不良 缺失率 寡核苷酸引物 Duchenne

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。

The authors used molecular biology techniques to amplify nine pairs of DNA sequences of the dystrophia gene using nine sets of oligonucleotide primers in two-step multiplex polymerase chain reaction (PCR). Nineteen Duchenne’s muscular dystrophy (DMD) and two Becker Typed muscular dystrophy (BMD) .Firstly, using 5 pairs of primers with high deletion rate, 8 cases were detected by deletion and 4 cases were amplified, The total PCR-multiplex PCR-negative rate of 9 pairs of primers accounted for 47.2% of the tested patients, indicating that about 90% of the patients with gene deletion could be detected.The experimental results suggest that two-step multiplex PCR can be used for gene diagnosis of DMD and / or BMD The simple, rapid method eliminates the hassle of using isotopes and is a good way to quickly diagnose DMD and / or BMD.