Based on three colorectal cancer cell lines with specified K-ras status, a sensitive PCR-RFLP assay was established detecting one K-ras mutant among 106 wild-type cells. Using this assay for tissues of 124 colorectal cancer patients, 59 tumor (46%) and 11 mucosa samples (9%) were found to harbor a K-ras mutation. When using the same scalpel for collecting tumor and mucosa tissues (group A), 18% of the patients had a matching K-ras mutation in both tissues, but this coincidence was seen in 3% of patients only, when separate scalpels were used (group B). Thus we conclude that the sampling technique used for collecting specimens is a major contributor to the detection of K-ras mutations in normal appearing mucosa when a highly sensitive detection technique is used.

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