Asymmetric real-time PCR detection of BRCA1 5382insC mutation by melting curve analysis in the LightCycler

Clin Chim Acta. 2008 Apr;390(1-2):141-4. doi: 10.1016/j.cca.2007.12.024. Epub 2008 Jan 17.

Abstract

Background: 5382insC BRCA1 frameshift mutation is a common founder mutation for many populations worldwide and a high-risk allele for the development of hereditary breast and/or ovarian cancer. Our goal was to develop a novel, reliable and rapid method for its detection.

Methods: We developed an asymmetric real-time PCR method with hybridization probes in the LightCycler. Genotyping was performed by melting curve analysis.

Results and conclusions: The developed method was in concordance with reference methods when tested in 85 peripheral blood and 107 tumor DNA samples from Greek breast and/or ovarian cancer patients. The described method proved to be simple, cost-effective, easy to perform and rapid enough for routine use as a screening method in high-risk families and especially in the Greek, Slavic and Jewish populations where 5382insC mutation is the most common BRCA1 mutation.

MeSH terms

  • BRCA1 Protein / genetics*
  • Base Sequence
  • Breast Neoplasms / genetics
  • DNA Primers
  • Female
  • Humans
  • Mutation*
  • Ovarian Neoplasms / genetics
  • Polymerase Chain Reaction / methods*

Substances

  • BRCA1 Protein
  • DNA Primers