Automated Detection of Prevalent Mutations in BRCA1 and BRCA2 Genes, Using a Fluorogenic PCR Allelic Discrimination Assay

摘要: ABSTRACT Mutations in the genes BRCA1 and BRCA2 account for 5%–10% of familial early onset breast cancer. Identification these mutations allows molecular diagnosis cancer susceptibility. A high through-put automated PCR allelic discrimination assay (ADA) was developed to detect prevalent genes. Two allele specific oligonucleotides (ASO) are directly used reaction, both which fluorescent reporter quencher dyes attached 5′ 3′ ends, respectively. During PCR, fluorescence is generated after cleavage annealed ASO by nuclease activity Taq polymerase. The wild-type BRCA sequence distinguished from mutant differential emission two different dyes. sensitivity ADA at level a single cell following nested PCR. Eighty-six patient samples can be analyzed each mutation 15-min post-PCR without need radioactivity, gel electrophoresis, or membrane bl...