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


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.
Kroupis
Christos
C
Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens 15771, Greece.
Christopoulos
Kostantinos
K
Devetzoglou
Myrto
M
Tsiagas
Ioannis
I
Lianidou
Evi S
ES
D019313:BRCA1 Protein / Q000235:genetics*; D001483:Base Sequence; D001943:Breast Neoplasms / Q000235:genetics; D017931:DNA Primers; D005260:Female; D006801:Humans; D009154:Mutation; D010051:Ovarian Neoplasms / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*
D019313:BRCA1 Protein; D017931:DNA Primers