Division of Genetics, Department of Biology, Faculty of Science, University of Isfahan, Isfahan, IR, Iran.
Division of Biotechnology, Department of Agricultural Biotechnology, Faculty of Agriculture, Isfahan University of Technology, Isfahan, IR, Iran.
Metab Brain Dis. 2018 Aug;33(4):1165-1173. doi: 10.1007/s11011-018-0210-x. Epub 2018 Apr 3.
In this study, we introduce a novel compound-primed multiplex ARMS PCR (CPMAP) for simultaneous detection of common PAH gene mutations. This approach was used successfully for simultaneous identification of six most common PAH gene mutations in 137 phenylketonuria patients in the Iranian population. A total of six normal and six mutant allele-specific primers and 4 common primers containing a tag sequence of 12 base pair at the 5'-end were designed and used in two separate optimized multiplex ARMS reactions followed by hot-start PCR. The products were separated and visualized on 3% agarose gel. The CPMAP genotyping data were completely in accordance with the direct sequencing results. The CPMAP suggests a reliable, economical and rapid method for simultaneous detection of PAH point mutations using conventional PCR, which could be applied for diagnosis of other gene mutations.
在这项研究中，我们介绍了一种新的复合引物多重扩增阻滞突变系统（CPMAP），用于同时检测常见的苯丙氨酸羟化酶基因突变。该方法成功地用于同时鉴定 137 名伊朗人群中苯丙酮尿症患者的六种最常见的 PAH 基因突变。总共设计了六个正常和六个突变等位基因特异性引物和 4 个通用引物，它们在两个单独的优化多重扩增阻滞突变系统反应后使用，接着是热启动 PCR。产物在 3%琼脂糖凝胶上分离和可视化。CPMAP 基因分型数据与直接测序结果完全一致。CPMAP 表明，使用常规 PCR 同时检测 PAH 点突变是一种可靠、经济和快速的方法，可用于诊断其他基因突变。
