Reliability of KRAS mutation testing in metastatic colorectal cancer patients across five laboratories

BMC Res Notes. 2012 Apr 25:5:196. doi: 10.1186/1756-0500-5-196.

Abstract

Background: Mutations in the KRAS gene are associated with poor response to epidermal growth factor receptor inhibitors used in the treatment of metastatic colorectal cancer. Factors influencing KRAS test results in tumor specimens include: tumor heterogeneity, sample handling, slide preparation, techniques for tumor enrichment, DNA preparation, assay design and sensitivity. We evaluated comparability and consistency of KRAS test results among five laboratories currently being used to determine KRAS mutation status of metastatic colorectal cancer specimens in a large, multi-center observational study.

Findings: Twenty formalin-fixed paraffin-embedded human colorectal cancer samples from colon resections previously tested for KRAS mutations were selected based on mutation status (6 wild type, 8 codon 12 mutations, and 6 codon 13 mutations). We found good agreement across laboratories despite differences in mutation detection methods. Eighteen of twenty samples (90%) were concordant across all five labs. Discordant results are likely not due to laboratory error, but instead to tumor heterogeneity, contamination of the tumor sample with normal tissue, or analytic factors affecting assay sensitivity.

Conclusions: Our results indicate commercial and academic laboratories provide reliable results for the common KRAS gene mutations at codons 12 and 13 when an adequate percentage of tumor cells is present in the sample.

Publication types

  • Multicenter Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / genetics*
  • Adenocarcinoma / secondary
  • Aged
  • Aged, 80 and over
  • Carcinoma / genetics*
  • Carcinoma / secondary
  • Clinical Laboratory Techniques / standards*
  • Codon
  • Colorectal Neoplasms / genetics*
  • Colorectal Neoplasms / pathology
  • DNA Mutational Analysis / standards*
  • Guidelines as Topic / standards
  • Humans
  • Middle Aged
  • Mutation*
  • Observer Variation
  • Paraffin Embedding
  • Predictive Value of Tests
  • Proto-Oncogene Proteins / genetics*
  • Proto-Oncogene Proteins p21(ras)
  • Quality Control
  • Real-Time Polymerase Chain Reaction / standards
  • Reproducibility of Results
  • Tissue Fixation
  • United States
  • ras Proteins / genetics*

Substances

  • Codon
  • KRAS protein, human
  • Proto-Oncogene Proteins
  • Proto-Oncogene Proteins p21(ras)
  • ras Proteins