Wang Yaping, Friedl Waltraut, Sengteller Marlies, Jungck Matthias, Filges Isabel, Propping Peter, Mangold Elisabeth
Institute of Human Genetics, University of Bonn, Bonn, Germany.
Hum Mutat. 2002 Mar;19(3):279-86. doi: 10.1002/humu.10042.
A method for detection of large genomic deletions in the MSH2 and MLH1 genes based on multiplex PCR and quantitative evaluation of PCR products is presented. All 35 exons of MSH2 and MLH1 were screened simultaneously in seven PCR reactions, each of them including primers for both genes. The method is reliable for uncovering large genomic deletions in patients suspected of HNPCC. With this method, six novel deletions were identified, two in MSH2: EX1_10del and EX1_16del (representing deletion of the entire MSH2 gene); and four in MLH1: EX1_10del in two unrelated patients, EX3_5del, and EX4del. The deletions were detected in 18 unrelated patients in whom no germline mutation had been identified by SSCP and DHPLC. These results indicate that our modified multiplex PCR assay is suited for the detection of large deletions both in the MSH2 and MLH1 gene and therefore represents an additional valuable tool for mutation screening in HNPCC families.
本文介绍了一种基于多重PCR和PCR产物定量评估来检测MSH2和MLH1基因大片段基因组缺失的方法。在七个PCR反应中同时筛选MSH2和MLH1的所有35个外显子，每个反应都包含两个基因的引物。该方法对于发现疑似遗传性非息肉病性结直肠癌（HNPCC）患者的大片段基因组缺失是可靠的。通过该方法，鉴定出六个新的缺失，其中两个在MSH2中：EX1_10del和EX1_16del（代表整个MSH2基因的缺失）；四个在MLH1中：两个无关患者中的EX1_10del、EX3_5del和EX4del。在18名无关患者中检测到这些缺失，这些患者通过单链构象多态性（SSCP）和变性高效液相色谱（DHPLC）未鉴定出种系突变。这些结果表明，我们改进的多重PCR检测方法适用于检测MSH2和MLH1基因中的大片段缺失，因此是HNPCC家系突变筛查的另一个有价值的工具。
