Detection of Point Mutations in Kirsten ras 2 Gene Using Locked Nucleic Acids Clamped PCR
AbstraktThe aim of the study was to examine diagnostic possibilities of LNA (locked nucleic acids) -clamped PCR for detection of somatic mutations in the K-ras gene in 133 samples of human malignant colorectal tumors. Selective real-time PCR amplification of mutant alleles of the K-ras gene (codon 12 and 13) revealed the presence of mutations in 45 samples of the tumors. The detection limit of the used method for K-ras mutant alleles was 0.03 ng. LNA-clamped PCR suppressing amplification of the wild-type alleles of the K-ras gene provides a very sensitive and specific detection of mutations present in DNA samples of colorectal carcinomas.