Fluorescence-based Detection of the CETP TaqIB Polymorphism: False Positives with the TaqMan-based Exonuclease Assay Attributable to a Previously Unknown Gene Variant

摘要: Background: Previous studies have shown an association between the Taq IB polymorphism of cholesteryl ester transfer protein ( CETP ) gene with plasma and HDL concentrations progression coronary artery disease (CAD). The aim present study was to determine performance two new fluorescence-based detection systems in analysis genotype. Methods: CAD patients (n = 150) known genotype, as determined by restriction fragment length (RFLP) analysis, were selected, including three groups 50 patients, carrying B1/1, B1/2, B2/2 genotypes, respectively. genotypes also analyzed allele-specific TaqMan PCR melting curve (LightCycler). In addition, DNA sequencing applied. Results: obtained fluorescence corresponded those RFLP exception heterozygous (B1/2), who misclassified homozygous B2 carriers system. Melting these samples demonstrated additional point at 59.1 °C, which found four for B1 allele. revealed a previously unknown C270T nucleotide exchange intron 1 gene, only nine base pairs from site. Conclusions: Determination system led misclassifications because gene. substitution detected LightCycler occurrence point. Our data indicate importance thorough evaluation before using them on routine basis.