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

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  • 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.

Publikováno

15.10.2007

Jak citovat

Beránek, M., Bureš, J., Šácha, M., Sákra, L., Rajman, M., Jandík, P., Rudolf, E., & Landt, O. (2007). Detection of Point Mutations in Kirsten ras 2 Gene Using Locked Nucleic Acids Clamped PCR. Chemické Listy, 101(9). Získáno z http://ww.chemicke-listy.cz/ojs3/index.php/chemicke-listy/article/view/1748

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