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

BMC Research Notes

C Sue Richards¹  Loic Le Marchand³  Dhananjay A Chitale⁵  Tia L Kauffman²  Alanna Kulchak Rahm⁴  Kellyan C Funk⁴  Monique A Johnson¹  Katrina AB Goddard²  Heather Spencer Feigelson⁴ 

[1] Molecular and Medical Genetics, Oregon Health & Science University, 3181 S.W. Sam Jackson Park Road, Portland, OR, 97239-3098, USA;Kaiser Permanente Northwest, Center for Health Research, 3800 N. Interstate Avenue, Portland, OR, 97227, USA;University of Hawaii Cancer Center, 677 Ala Moana Boulevard, Suite 901, Honolulu, HI, 96813, USA;Institute for Health Research Legacy Highlands, Kaiser Permanente Colorado, Suite 300, P.O. Box 378066, Denver, CO, 80237-8066, USA;Department of Pathology and Laboratory Medicine, Henry Ford Hospital, 2799 W. Grand Blvd., Detroit, MI, 48202, USA

关键词: Laboratory error;    EGFR;    Colorectal cancer;    KRAS;   

Others  :  1166488
DOI  :  10.1186/1756-0500-5-196

 received in 2012-01-17, accepted in 2012-04-25,  发布年份 2012

【 摘 要 】

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.

【 授权许可】

   
2012 Feigelson et al.; licensee BioMed Central Ltd.

【 参考文献 】

• [1]Dahabreh IJ, Terasawa T, Castaldi PJ, Trikalinos TA: Systematic review: Anti-epidermal growth factor receptor treatment effect modification by KRAS mutations in advanced colorectal cancer. Ann Intern Med 2011, 154:37-49.
• [2]Allegra CJ, Jessup JM, Somerfield MR, Hamilton SR, Hammond EH, Hayes DF, et al.: American Society of Clinical Oncology provisional clinical opinion: testing for KRAS gene mutations in patients with metastatic colorectal carcinoma to predict response to anti-epidermal growth factor receptor monoclonal antibody therapy. J Clin Oncol 2009, 27:2091-2096.
• [3]Franklin WA, Haney J, Sugita M, Bemis L, Jimeno A, Messersmith WA: KRAS mutation: comparison of testing methods and tissue sampling techniques in colon cancer. J Mol Diagn 2010, 12:43-50.
• [4]Holdhoff M, Schmidt K, Donehower R, Diaz LA: Analysis of circulating tumor DNA to confirm somatic KRAS mutations. J Natl Cancer Inst 2009, 101:1284-1285.
• [5]Jimeno A, Messersmith WA, Hirsch FR, Franklin WA, Eckhardt SG: KRAS mutations and sensitivity to epidermal growth factor receptor inhibitors in colorectal cancer: practical application of patient selection. J Clin Oncol 2009, 27:1130-1136.
• [6]Loupakis F, Ruzzo A, Cremolini C, Vincenzi B, Salvatore L, Santini D, et al.: KRAS codon 61, 146 and BRAF mutations predict resistance to cetuximab plus irinotecan in KRAS codon 12 and 13 wild-type metastatic colorectal cancer. Br J Cancer 2009, 101:715-721.
• [7]Janakiraman M, Vakiani E, Zeng Z, Pratilas CA, Taylor BS, Chitale D, et al.: Genomic and biological characterization of exon 4 KRAS mutations in human cancer. Cancer Res 2010, 70:5901-5911.
• [8]Carotenuto P, Roma C, Rachiglio AM, Tatangelo F, Pinto C, Ciardiello F, et al.: Detection of KRAS mutations in colorectal carcinoma patients with an integrated PCR/sequencing and real-time PCR approach. Pharmacogenomics 2010, 11:1169-1179.
• [9]Whitehall V, Tran K, Umapathy A, Grieu F, Hewitt C, Evans TJ, et al.: A multicenter blinded study to evaluate KRAS mutation testing methodologies in the clinical setting. J Mol Diagn 2009, 11:543-552.
• [10]Weichert W, Schewe C, Lehmann A, Sers C, Denkert C, Budczies J, et al.: KRAS genotyping of paraffin-embedded colorectal cancer tissue in routine diagnostics: comparison of methods and impact of histology. J Mol Diagn 2010, 12:35-42.
• [11]Tsiatis AC, Norris-Kirby A, Rich RG, Hafez MJ, Gocke CD, Eshleman JR, et al.: Comparison of Sanger sequencing, pyrosequencing, and melting curve analysis for the detection of KRAS mutations: Diagnostic and clinical implications. J Mol Diagn 2010, 12:1-8.
• [12]Angulo B, Garcia-Garcia E, Martinez R, Suarez-Gauthier A, Conde E, Hidalgo M, et al.: A commercial real-time PCR kit provides greater sensitivity than direct sequencing to detect KRAS mutations: a morphology-based approach in colorectal carcinoma. J Mol Diagn 2010, 12:292-299.
• [13]Oliner K, Juan T, Suggs S, Wolf M, Sarosi I, Freeman DJ, et al.: A comparability study of 5 commercial KRAS tests. Diagn Pathol 2010, 5:23. BioMed Central Full Text