KRAS mutation: Comparison of testing methods and tissue sampling techniques in colon cancer

Wilbur A. Franklin, Jerry Haney, Michio Sugita, Lynne Bemis, Antonio Jimeno, Wells A. Messersmith

Research output: Contribution to journalArticlepeer-review

104 Scopus citations


Treatment of colon carcinoma with the anti-epidermal growth factor receptor antibody Cetuximab is reported to be ineffective in KRAS-mutant tumors. Mutation testing techniques have therefore become an urgent concern. We have compared three methods for detecting KRAS mutations in 59 cases of colon carcinoma: 1) high resolution melting, 2) the amplification refractory mutation system using a bifunctional self-probing primer (ARMS/Scorpion, ARMS/S), and 3) direct sequencing. We also evaluated the effects of the methods of sectioning and coring of paraffin blocks to obtain tumor DNA on assay sensitivity and specificity. The most sensitive and specific combination of block sampling and mutational analysis was ARMS/S performed on DNA derived from 1-mm paraffin cores. This combination of tissue sampling and testing method detected KRAS mutations in 46% of colon tumors. Four samples were positive by ARMS/S, but initially negative by direct sequencing. Cloned DNA samples were retested by direct sequencing, and in all four cases KRAS mutations were identified in the DNA. In six cases, high resolution melting abnormalities could not be confirmed as specific mutations either by ARMS/S or direct sequencing. We conclude that coring of the paraffin blocks and testing by ARMS/S is a sensitive, specific, and efficient method for KRAS testing.

Original languageEnglish (US)
Pages (from-to)43-50
Number of pages8
JournalJournal of Molecular Diagnostics
Issue number1
StatePublished - Jan 2010

Bibliographical note

Funding Information:
Supported by NIH NCI Cancer Center grant P30-CA046934 and NIH NCI Early Detection Research Network Grant U01-CA85070 .


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