A highly accurate, low cost test forBRCA1 mutations
- Nathalie J van Orsouwa,
- Rahul K Dhandab,
- Youssef Elhajic,
- Steven A Narodc,
- Frederick P Lid,
- Charis Enge,
- Jan Vijga
- aInstitute for Drug Development, Cancer Therapy and Research Center, 8122 Datapoint Drive, Suite 700, San Antonio, TX 78229, USA, bMosaic Technologies Inc, 1106 Commonwealth Avenue, Boston, MA 02215, USA, cOntario Cancer Institute, Princess Margaret Hospital, 610 University Avenue, Toronto, Ontario, Canada M5G 2M9, dProgram in Population Sciences, Dana-Farber Cancer Institute and Harvard Medical School, 44 Binney Street, Boston, MA 02115, USA, eClinical Cancer Genetics and Human Cancer Genetics Programs, Ohio State University Comprehensive Cancer Center, 420 W 12th Avenue, 690c MRF, Columbus, OH 43210, USA
- Dr van Orsouw.nvorsouw{at}saci.org
- Revised 2 June 1999
- Accepted 21 June 1999
Abstract
The hereditary breast and ovarian cancer syndrome is associated with a high frequency of BRCA1mutations. However, the widespread use ofBRCA1 testing has been limited to date by three principal concerns: the fear of loss of health and life insurance, the uncertain clinical value of a positive test result, and the current lack of an inexpensive and sensitive screening test forBRCA1 mutations. We have developed an inexpensive system for gene mutational scanning, based on a combination of extensive multiplex PCR amplification and two dimensional electrophoresis. The efficiency of this system, as a screening test forBRCA1 mutations, was evaluated in a panel of 60 samples from high risk women, 14 of which contained a previously identified mutation in BRCA1. All 14 mutations were identified, as well as an additional five that had previously escaped detection. In addition to the 19 mutations, a total of 15 different polymorphic variants were scored, most of which were recurring. All were confirmed by nucleotide sequencing. The cost of screening per sample was calculated to be approximately US$70 for the manual technique used in this study, and may be reduced to approximately US$10 with the introduction of commercially available PCR robotics and fluorescent imaging. Implementation of this method of mutation screening in the research and clinical setting should permit rapid accrual of quantitative data on genotype-phenotype associations for the evaluation of diagnostic testing.
