Portal de Periódicos da CAPES

A rapid fluorescent multiplexed‐PCR analysis (FMPA) for founder mutations in the BRCA 1 and BRCA 2 genes

2000; Wiley; Volume: 57; Issue: 3 Linguagem: Inglês

10.1034/j.1399-0004.2000.570307.x

1399-0004

Graciela Kuperstein, William D. Foulkes, Parviz Ghadirian, Jalil Hakimi, Steven A. Narod,

DNA Repair Mechanisms

Mutations of the BRCA 1 and BRCA 2 genes account for approximately 80% of hereditary breast/ovarian cancer families, but the size of these two genes makes mutation analysis time‐consuming and technically challenging. In some populations such as the Ashkenazi Jewish and the French‐Canadian, a small number of recurrent founder mutations account for the majority of mutations in cancer families. We have therefore developed two rapid genetic screening tests, which allow us to detect three frequent frameshift mutations in the Ashkenazi Jewish population and five frameshift mutations in the French‐Canadian population. These fluorescent non‐radioactive methods permit the simultaneous detection of multiple mutations by generating multiplexed PCR‐amplified gene fragments, and by discriminating these on the basis of their size in a denaturing polyacrylamide gel. Using these methods, we were able to correctly identify all mutants in a blinded analysis of 276 DNA samples, including 30 derived from paraffin‐embedded tumor samples and 10 from buccal‐cell brushes, with no false positive or false negative results. These techniques designed for the direct detection of recurrent mutations in the BRCA 1 and BRCA 2 genes, have the advantages of being efficient, sensitive, cost‐effective, and are applicable to large scale screening for epidemiologic studies.