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KRAS Mutation Test in Korean Patients with Colorectal Carcinomas: A Methodological Comparison between Sanger Sequencing and a Real-Time PCR-Based Assay

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Background: Mutations in the KRAS gene have been identified in approximately 50% of colorectal cancers (CRCs). KRAS mutations are well established biomarkers in anti–epidermal growth factor receptor therapy. Therefore, assessment of KRAS mutations is needed in CRC patients to ensure appropriate treatment. Methods: We compared the analytical performance of the cobas test to Sanger sequencing in 264 CRC cases. In addition, discordant specimens were evaluated by 454 pyrosequencing. Results: KRAS mutations for codons 12/13 were detected in 43.2% of cases (114/264) by Sanger sequencing. Of 257 evaluable specimens for comparison, KRAS mutations were detected in 112 cases (43.6%) by Sanger sequencing and 118 cases (45.9%) by the cobas test. Concordance between the cobas test and Sanger sequencing for each lot was 93.8% positive percent agreement (PPA) and 91.0% negative percent agreement (NPA) for codons 12/13. Results from the cobas test and Sanger sequencing were discordant for 20 cases (7.8%). Twenty discrepant cases were subsequently subjected to 454 pyrosequencing. After comprehensive analysis of the results from combined Sanger sequencing–454 pyrosequencing and the cobas test, PPA was 97.5% and NPA was 100%. Conclusions: The cobas test is an accurate and sensitive test for detecting KRAS-activating mutations and has analytical power equivalent to Sanger sequencing. Prescreening using the cobas test with subsequent application of Sanger sequencing is the best strategy for routine detection of KRAS mutations in CRC.

Abstract

Background: Mutations in the KRAS gene have been identified in approximately 50% of colorectal cancers (CRCs). KRAS mutations are well established biomarkers in anti–epidermal growth factor receptor therapy. Therefore, assessment of KRAS mutations is needed in CRC patients to ensure appropriate treatment. Methods: We compared the analytical performance of the cobas test to Sanger sequencing in 264 CRC cases. In addition, discordant specimens were evaluated by 454 pyrosequencing. Results: KRAS mutations for codons 12/13 were detected in 43.2% of cases (114/264) by Sanger sequencing. Of 257 evaluable specimens for comparison, KRAS mutations were detected in 112 cases (43.6%) by Sanger sequencing and 118 cases (45.9%) by the cobas test. Concordance between the cobas test and Sanger sequencing for each lot was 93.8% positive percent agreement (PPA) and 91.0% negative percent agreement (NPA) for codons 12/13. Results from the cobas test and Sanger sequencing were discordant for 20 cases (7.8%). Twenty discrepant cases were subsequently subjected to 454 pyrosequencing. After comprehensive analysis of the results from combined Sanger sequencing–454 pyrosequencing and the cobas test, PPA was 97.5% and NPA was 100%. Conclusions: The cobas test is an accurate and sensitive test for detecting KRAS-activating mutations and has analytical power equivalent to Sanger sequencing. Prescreening using the cobas test with subsequent application of Sanger sequencing is the best strategy for routine detection of KRAS mutations in CRC.

참고문헌 (38)

  1. Malumbres M, "RAS oncogenes: the first 30 years", Nat Rev Cancer, 3, pp.459~465 (2003)
  2. Bos JL, "Ras oncogenes in human cancer : a review", Cancer Res, 49, pp.4682~4689 (1989)
  3. Lee S, "Analytical performance of a PCR assay for the detection of KRAS mutations (codons 12", 13 and 61) in formalin-fixed paraffin-embedded tissue samples of colorectal carcinoma, 2012, pp.460, 141~149
  4. Venook AP, "Epidermal growth factor receptor-targeted treatment for advanced colorectal carcinoma", Cancer, 103, pp.2435~2446 (2005)
  5. Dahabreh IJ, "Systematic review: anti-epidermal growth factor receptor treatment effect modification by KRAS mutations in advanced colorectal cancer", Ann Intern Med, 154, pp.37~49 (2011)
  6. Querings S, "Benchmarking of mutation diagnostics in clinical lung cancer specimens", PLoS One, 6, pp.e19601~ (2011)
  7. Kim SY, "Syndecan-1 expression is associated with tumor size and EGFR expression in colorectal carcinoma: a clinicopathological study of 230 cases", Int J Med Sci, 12, pp.92~99 (2015)
  8. Harlé A, "Comparison of COBAS 4800 KRAS, TaqMan PCR and high resolution melting PCR assays for the detection of KRAS somatic mutations in formalin-fixed paraffin embedded colorectal carcinomas", Virchows Arch, 462, pp.329~335 (2013)
  9. Sharma A, "Novel approach for clinical validation of the cobas KRAS mutation test in advanced colorectal cancer", Mol Diagn Ther, 20, pp.231~240 (2016)
  10. Gonzalez de Castro D, "A comparison of three methods for detecting KRAS mutations in formalin-fixed colorectal cancer specimens", Br J Cancer, 107, pp.345~351 (2012)
  11. Amado RG, "Wild-type KRAS is required for panitumumab efficacy in patients with metastatic colorectal cancer", J Clin Oncol, 26, pp.1626~1634 (2008)
  12. Van Cutsem E, "Cetuximab and chemotherapy as initial treatment for metastatic colorectal cancer", N Engl J Med, 360, pp.1408~1417 (2009)
  13. Bokemeyer C, "Fluorouracil, leucovorin, and oxaliplatin with and without cetuximab in the firstline treatment of metastatic colorectal cancer", J Clin Oncol, 27, pp.663~671 (2009)
  14. Douillard JY, "Randomized, phase III trial of panitumumab with infusional fluorouracil, leucovorin, and oxaliplatin (FOLFOX4) versus FOLFOX4 alone as first-line treatment in patients with previously untreated metastatic colorectal cancer: the PRIME study", J Clin Oncol, 28, pp.4697~4705 (2010)
  15. Malapelle U, "KRAS mutation detection by high-resolution melting analysis significantly predicts clinical benefit of cetuximab in metastatic colorectal cancer", Br J Cancer, 107, pp.626~631 (2012)
  16. Malapelle U, "KRAS testing in metastatic colorectal carcinoma: challenges, controversies, breakthroughs and beyond", J Clin Pathol, 67, pp.1~9 (2014)
  17. Angulo B, "A new generation of companion diagnostics: cobas BRAF, KRAS and EGFR mutation detection tests", Expert Rev Mol Diagn, 14, pp.517~524 (2014)
  18. Do H, "Sequence artifacts in DNA from formalin-fixed tissues: causes and strategies for minimization", Clin Chem, 61, pp.64~71 (2015)
  19. Sah S, "Functional DNA quantification guides accurate next-generation sequencing mutation detection in formalin-fixed, paraffin-embedded tumor biopsies", Genome Med, 5, pp.77~ (2013)
  20. Tsiatis AC, "Comparison of Sanger sequencing, pyrosequencing, and melting curve analysis for the detection of KRAS mutations: diagnostic and clinical implications", J Mol Diagn, 12, pp.425~432 (2010)
  21. Costello M, "Discovery and characterization of artifactual mutations in deep coverage targeted capture sequencing data due to oxidative DNA damage during sample preparation", Nucleic Acids Res, 41, pp.e67~ (2013)
  22. Kinde I, "Detection and quantification of rare mutations with massively parallel sequencing", Proc Natl Acad Sci U S A, 108, pp.9530~9535 (2011)
  23. Xuan J, "Next-generation sequencing in the clinic: promises and challenges", Cancer Lett, 340, pp.284~295 (2013)
  24. Li H, "Mapping short DNA sequencing reads and calling variants using mapping quality scores", Genome Res, 18, pp.1851~1858 (2008)
  25. Quach N, "In vitro mutation artifacts after formalin fixation and error prone translesion synthesis during PCR", BMC Clin Pathol, 4, pp.1~ (2004)
  26. Wong SQ, "Sequence artefacts in a prospective series of formalin-fixed tumours tested for mutations in hotspot regions by massively parallel sequencing", BMC Med Genomics, 7, pp.23~ (2014)
  27. De Roock W, "Association of KRAS p.G13D mutation with outcome in patients with chemotherapy-refractory metastatic colorectal cancer treated with cetuximab", JAMA, 304, pp.1812~1820 (2010)
  28. Benvenuti S, "Oncogenic activation of the RAS", RAF signaling pathway impairs the response of metastatic colorectal cancers to anti-epidermal growth factor receptor antibody therapies, 2007, pp.67, 2643~2648
  29. Misale S, "Emergence of KRAS mutations and acquired resistance to anti-EGFR therapy in colorectal cancer", Nature, 486, pp.532~536 (2012)
  30. Morris VK, "Clinicopathologic characteristics and gene expression analyses of non-KRAS 12", 13, RASmutated metastatic colorectal cancer, 2014, pp.25, 2008~2014
  31. Loupakis F, "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, 101, pp.715~721 (2009)
  32. De Roock W, "Effects of KRAS, BRAF, NRAS, and PIK3CA mutations on the efficacy of cetuximab plus chemotherapy in chemotherapy-refractory metastatic colorectal cancer: a retrospective consortium analysis", Lancet Oncol, 11, pp.753~762 (2010)
  33. Gao J, "Comparative analysis of dideoxy sequencing, the KRAS StripAssay and pyrosequencing for detection of KRAS mutation", World J Gastroenterol, 16, pp.4858~4864 (2010)
  34. Angulo B, "A commercial realtime PCR kit provides greater sensitivity than direct sequencing to detect KRAS mutations: a morphology-based approach in colorectal carcinoma", J Mol Diagn, 12, pp.292~299 (2010)
  35. Whitehall V, "A multicenter blinded study to evaluate KRAS mutation testing methodologies in the clinical setting", J Mol Diagn, 11, pp.543~552 (2009)
  36. project-part 1), "J Thorac Oncol", 2011, 1006~1015
  37. Richman SD, "KRAS and BRAF mutations in advanced colorectal cancer are associated with poor prognosis but do not preclude benefit from oxaliplatin or irinotecan: results from the MRC FOCUS trial", J Clin Oncol, 27, pp.5931~5937 (2009)
  38. Gaedcke J, "KRAS and BRAF mutations in patients with rectal cancer treated with preoperative chemoradiotherapy", Radiother Oncol, 94, pp.76~81 (2010)

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