DHPLC mutation analysis of the hereditary nonpolyposis colon cancer (HNPCC) genes hMLH1 and hMSH2

Denaturing high-performance liquid chromatography (DHPLC) is an efficient m ethod for detection of mutations involving a single or few numbers of nucle otides, and it has been successfully used for mutation detection in disease -related genes. Colorectal cancer is one of the most common cancers, and mu tations in the genes for hereditary nonpolyposis colon cancer (HNPCC), hMLH 1 and hMSH2, also involve mainly point mutations. Sequence analysis is supp osed to be a screening method with high sensitivity; however, it is time-co nsuming and expensive. We therefore decided to test sensitivity and reprodu cibility of DHPLC for 71 sequence variants in hMLH1 and hMSH2 initially fou nd by sequence analysis in DNA samples of German HNPCC patients. DHPLC cond itions of the PCR products were based on the melting pattern of the wild-ty pe sequence of the corresponding PCR fragments. All but one of the 71 mutat ions was detected using DHPLC (sensitivity of 97%). Running time pet sample averaged only 7 min, and the system is highly automated. Thus DHPLC is a r apid and sensitive method for the detection of hMLH1 and hMSH2 sequence var iants. (C) 2001 published by Elsevier Science B.V.