PCR技术在疾病基因检测中的应用.pdfVIP

重复引物 PCR 技术在超大片段动态突变疾病基因检测 中的应用 1 2 陈晟 ，吴志英 1. 福建医科大学附属第一医院神经内科，福州350005 ； 2. 复旦大学附属华山医院神经内科，上海 200040 摘要：动态突变疾病是指基因编码区或非编码区发生核苷酸重复序列异常扩增所导致的一类遗传性疾病。发生 于非翻译区的动态突变常常伴有超大片段重复序列，应用普通PCR 法无法对该片段进行扩增，而传统的Southern blot 等技术费时费力，无法应用于临床基因诊断。在此背景下，重复引物PCR 技术应运而生，并随着应用范围 的扩大而逐渐改进，适用于强直性肌营养不良症、Friedreich 共济失调、脊髓小脑性共济失调 10 型及C9orf72 基 因突变引起的额颞叶痴呆或肌萎缩侧索硬化等遗传性动态突变疾病的临床基因检测。文章简要介绍了重复引物 PCR 技术的原理，着重阐述了重复引物PCR 技术在相关超大片段动态突变疾病临床基因检测中的应用进展。 关键词：重复引物PCR 技术；超大片段重复序列；基因诊断；动态突变疾病 Advances on repeat-primed PCR assay for the genetic diagnosis of dynamic mutation diseases with large pathogenic expansions 1 2 Sheng Chen , Zhiying Wu 1. Department of Neurology, First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, China; 2. Department of Neurology, Huashan Hospital, Fudan University, Shanghai 200040, China Abstract: Dynamic mutation diseases are genetic diseases caused by unstable repeat expansions in coding or noncoding regions. The unstable repeat expansions located in the noncoding region usually accompany large expansions which are difficult to amplify using the standard PCR assay. Traditional detection methods, including Southern blot, are usually time-consuming and labor-wasting. A new method called fluorescent repeat-primed PCR assay was brought into clinical applications. With the development of genetic diagnoses for dynamic mutation diseases, such as myotonic dystrophy, Friedreich’s ataxia, SCA10, and amyotrophic lateral sclerosis or frontotemporal dementia caused by C9orf72 mutations, the clinical use of repeat-primed PCR assay has been broadened. Here, we review the principle of repeat-primed PCR assay and its advances on the genetic diagnoses of related dynamic mutation diseases with large pathogenic expansions. Keywords: repeat-primed PCR assay; large pathogenic expansions; genetic diagnosis; dynamic mutation disease 动态突变疾病是指