Simple and Rapid Detection of BRCA1 and BRCA2 Mutations by Multiplex Mutagenically Separated PCR

摘要

BRCA1 and BRCA2 are tumor suppressor genes that are inactivated during neoplastic development (1)(2). Germline mutations of the two genes are transmitted in the autosomal dominant fashion and predispose carriers to the development of ovarian and/or breast cancers (3)(4). Mutations in BRCA1 are present in approximately one-half of the early-onset breast cancer families and 80% of the early-onset breast and ovarian cancer families (5), whereas BRCA2 mutations are believed to account for a comparable percentage of inherited breast cancer cases (6). Women with germline mutations in BRCA1 have a lifetime risk of 85% and up to 50% for breast and ovarian cancers, respectively.In individuals of Ashkenazi Jewish background, mutations such as 185delAG and 5382insC in BRCA1 , and 6174delT in BRCA2 are present in higher frequencies than other mutations because of founder effects (6)(7)(8). In one study, the three mutations accounted for 62% of Ashkenazi patients with ovarian and/or breast cancer (7). The high frequency of these mutations in the BRCA genes indicates their potential role in identifying individuals at risk.Many methods have been reported for the study of BRCA mutations, including allele-specific oligonucleotide hybridization (8)(9), allele-specific PCR (10), PCR-mediated site-directed mutagenesis (11)(12), heteroduplex analysis (HDA) (13)(14)(15), single-strand conformation polymorphism (14)(16), and the protein truncation test (14)(15). Methods such as HDA, single-strand conformation polymorphism, and the protein …