Sensitive and specific detection of K-ras mutations in colon tumors by short oligonucleotide mass analysis

Matilde E. Lleonart, Santiago Ramon Y. Cajal, John D. Groopman, Marlin D. Friesen

Research output: Contribution to journalArticle

Abstract

Short oligonucleotide mass analysis (SOMA) is a technique by which small sequences of mutated and wild-Type DNA, produced by PCR amplification and restriction digestion, are characterized by HPLC-electrospray ionization tandem mass spectrometry. We have adapted the method to specifically detect two common point mutations at codon 12 of the c-K-ras gene. Mutations in DNA from 121 colon tumor samples were identified by SOMA and validated by comparison with sequencing. SOMA correctly identified 26 samples containing the 12GAT mutation and four samples containing the 12AGT mutation. Sequencing did not reveal mutant DNA in three samples out of the 26 samples shown by SOMA to contain the 12GAT mutation. In these three samples, the presence of mutant DNA was con-firmed by SOMA analysis after selective PCR ampli-fication in the presence of BstN1 restriction enzyme. Additional mutations in codons 12 and 13 were revealed by sequencing in 24 additional samples, and their presence did not interfere with the correct identification of G to A or G to T mutations in codon 12. These results provide the basis for a sensitive and specific method to detect c-K-ras codon 12-mutated DNA at levels below 10-12% of wild-Type DNA.

Original languageEnglish (US)
Article numbere53
JournalNucleic acids research
Volume32
Issue number5
DOIs
StatePublished - Mar 1 2004

ASJC Scopus subject areas

  • Genetics

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