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Detection of K-ras mutations in stools of patients with colorectal cancer by mutant-enriched PCR

✍ Scribed by Peter Nollau; Carsten Moser; Günter Weinland; Christoph Wagener

Publisher
John Wiley and Sons
Year
1996
Tongue
French
Weight
639 KB
Volume
66
Category
Article
ISSN
0020-7136

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✦ Synopsis

Mutant-enriched PCR was applied to the detection of mutations at codons I 2 and I 3 of K-ras genes in the stools of patients with colorectal cancer. Mutations were analyzed in stool samples obtained prior to surgery. Resected tumor specimens were screened for K-ras mutations by PCR-mediated RFLP analysis. Using normal stool samples, assay conditions were adjusted to optimal sensitivity and specificity. The following specimens were included in the stu* 16 stool samples corresponding to carcinomas in which K-ras mutations had bean identified; 7 randomly selected stool samples corresponding to carcinomas which were negative for K-ras mutations; I stool sample from a patient with non-Hodgkin's lymphoma. In 13 of the 16 stool samples (81%) corresponding to tumors in which K-ras mutations had been identified previously, K-ras mutations were detected. In 2 of the 7 stool samples corresponding to tumors in which K-ras mutations had not been detected by previous PCR-mediated RFLP analysis, K-ras mutations were also present Reanalyses of the tumors corresponding to these 2 positive stool samples by mutant-enriched PCR revealed a K-ras mutation in one of the tumors. The stool and tumor of the patient with non-Hodgkin's lymphoma were negative for K-ras mutations. DNA sequence analysis revealed that, for each of the K-ras mutations identified in stool samples, identical base substitutions were present in the corresponding tumor tissue.

The results indicate that tumor cells harboring K-ras mutations can be detected in the stools of patients with colorectal cancer by mutant-enriched PCR with high sensitivity and specificity. Because of the simplicity of the technique, it may be suitable for screening of stool samples for mutations of the K-ros gene.

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