Detection of Point Mutations in Kirsten ras 2 Gene Using Locked Nucleic Acids Clamped PCR

  • M. Beránek Department of clinical biochemistry and diagnostics, Faculty of Medicine and Faculty Hospital Hradec Králové,
  • J. Bureš Department of internal medicine, Faculty of Medicine and Faculty Hospital Hradec Králové,
  • M. Šácha Department of surgery, Regional Hospital Pardubice,
  • L. Sákra Department of surgery, Regional Hospital Pardubice,
  • M. Rajman Department of surgery, Regional Hospital Pardubice,
  • P. Jandík Department of surgery, Faculty of Medicine and Faculty Hospital Hradec Králové,
  • E. Rudolf Department of medical biology and genetics, Faculty of Medicine and Faculty Hospital Hradec Králové,
  • O. Landt TIB MOLBIOL, Berlin, Germany

Abstrakt

The 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.
Publikované
2007-10-15
Sekce
Články