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KRAS assay selection: sensitivity and accuracy in clinical application

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Abstract

KRAS mutation status is routinely tested before the administration of chemotherapeutic agents that target EGFR such as cetuximab and panitumumab. Various commercial assays to analyze KRAS mutational status are currently certified for in vitro diagnostic use. However, the differences in the operational characteristics of the different assays have not been investigated. Since different assays could be different in specificity, sensitivity and precision, studies to clarify the best option for an adequate determination of KRAS mutation status are necessary. Until a gold standard for testing KRAS mutation status is established, the treating physician must interpret the results of any given assay keeping this limitation in mind.

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References

  1. Jonker DJ, O’Callaghan CJ, Karapetis CS, Zalcberg JR, Tu D, Au HJ, Berry SR, Krahn M, Price T, Simes RJ et al (2007) Cetuximab for the treatment of colorectal cancer. N Engl J Med 357(20):2040–2048

    Article  PubMed  CAS  Google Scholar 

  2. Van Cutsem E, Siena S, Humblet Y, Canon JL, Maurel J, Bajetta E, Neyns B, Kotasek D, Santoro A, Scheithauer W et al (2008) An open-label, single-arm study assessing safety and efficacy of panitumumab in patients with metastatic colorectal cancer refractory to standard chemotherapy. Ann Oncol 19(1):92–98

    Article  PubMed  Google Scholar 

  3. Malumbres M, Barbacid M (2003) RAS oncogenes: the first 30 years. Nat Rev 3(6):459–465

    Article  CAS  Google Scholar 

  4. Jancik S, Drabek J, Radzioch D, Hajduch M (2010) Clinical relevance of KRAS in human cancers. J Biomed Biotechnol 2010:150960

    Article  PubMed  Google Scholar 

  5. Schubbert S, Shannon K, Bollag G (2007) Hyperactive Ras in developmental disorders and cancer. Nat Rev 7(4):295–308

    Article  CAS  Google Scholar 

  6. Amado RG, Wolf M, Peeters M, Van Cutsem E, Siena S, Freeman DJ, Juan T, Sikorski R, Suggs S, Radinsky R et al (2008) Wild-type KRAS is required for panitumumab efficacy in patients with metastatic colorectal cancer. J Clin Oncol 26(10):1626–1634

    Article  PubMed  CAS  Google Scholar 

  7. Herreros-Villanueva M, Rodrigo M, Claver M, Muniz P, Lastra E, Garcia-Giron C, Coma del Corral MJ (2010) KRAS, BRAF, EGFR and HER2 gene status in a Spanish population of colorectal cancer. Mol Biol Rep 38(2):1315–1320

    Article  PubMed  Google Scholar 

  8. Venook AP (2005) Epidermal growth factor receptor-targeted treatment for advanced colorectal carcinoma. Cancer 103(12):2435–2446

    Article  PubMed  CAS  Google Scholar 

  9. Kohne CH (2010) How to integrate molecular targeted agents in the continuum of care. Ann Oncol 21(Suppl 7):134–139

    Google Scholar 

  10. Richman SD, Seymour MT, Chambers P, Elliott F, Daly CL, Meade AM, Taylor G, Barrett JH, Quirke P (2009) 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(35):5931–5937

    Article  PubMed  CAS  Google Scholar 

  11. de Bruijn MT, Raats DA, Hoogwater FJ, van Houdt WJ, Cameron K, Medema JP, Borel Rinkes IH, Kranenburg O (2010) Oncogenic KRAS sensitises colorectal tumour cells to chemotherapy by p53-dependent induction of Noxa. Br J Cancer 102(8):1254–1264

    Article  PubMed  Google Scholar 

  12. Ogino S, Kawasaki T, Brahmandam M, Yan L, Cantor M, Namgyal C, Mino-Kenudson M, Lauwers GY, Loda M, Fuchs CS (2005) Sensitive sequencing method for KRAS mutation detection by Pyrosequencing. J Mol Diagn 7(3):413–421

    Article  PubMed  CAS  Google Scholar 

  13. Marchetti A, Buttitta F, Pellegrini S, Chella A, Bertacca G, Filardo A, Tognoni V, Ferreli F, Signorini E, Angeletti CA et al (1996) Bronchioloalveolar lung carcinomas: K-ras mutations are constant events in the mucinous subtype. J Pathol 179(3):254–259

    Article  PubMed  CAS  Google Scholar 

  14. Do H, Krypuy M, Mitchell PL, Fox SB, Dobrovic A (2008) High resolution melting analysis for rapid and sensitive EGFR and KRAS mutation detection in formalin fixed paraffin embedded biopsies. BMC Cancer 8:142–156

    Article  PubMed  Google Scholar 

  15. Lleonart ME, Ramon y Cajal S, Groopman JD, Friesen MD (2004) Sensitive and specific detection of K-ras mutations in colon tumors by short oligonucleotide mass analysis. Nucleic Acids Res 32(5):e53

    Article  PubMed  Google Scholar 

  16. Lilleberg SL, Durocher J, Sanders C, Walters K, Culver K (2004) High sensitivity scanning of colorectal tumors and matched plasma DNA for mutations in APC, TP53, K-RAS, and BRAF genes with a novel DHPLC fluorescence detection platform. Ann NY Acad Sci 1022:250–256

    Article  PubMed  CAS  Google Scholar 

  17. Ausch C, Buxhofer-Ausch V, Oberkanins C, Holzer B, Minai-Pour M, Jahn S, Dandachi N, Zeillinger R, Kriegshauser G (2009) Sensitive detection of KRAS mutations in archived formalin-fixed paraffin-embedded tissue using mutant-enriched PCR and reverse-hybridization. J Mol Diagn 11(6):508–513

    Article  PubMed  CAS  Google Scholar 

  18. Kitano S, Nakayama M, Yamane A, Tsukahara Y, Amano M (2011) Detection of DNA mutations by fluorescence resonance energy transfer-based preferential homoduplex formation assay. Anal Biochem 408(2):197–205

    Article  PubMed  CAS  Google Scholar 

  19. Nollau P, Wagener C (1997) Methods for detection of point mutations: performance and quality assessment. the IFCC Scientific Division, Committee on Molecular Biology Techniques. J Int Fed Clin Chem 9(4):162–170

    PubMed  CAS  Google Scholar 

  20. Haliassos A, Chomel JC, Grandjouan S, Kruh J, Kaplan JC, Kitzis A (1989) Detection of minority point mutations by modified PCR technique: a new approach for a sensitive diagnosis of tumor-progression markers. Nucleic Acids Res 17(20):8093–8099

    Article  PubMed  CAS  Google Scholar 

  21. Jiang W, Kahn SM, Guillem JG, Lu SH, Weinstein IB (1989) Rapid detection of ras oncogenes in human tumors: applications to colon, esophageal, and gastric cancer. Oncogene 4(7):923–928

    PubMed  CAS  Google Scholar 

  22. Levi S, Urbano-Ispizua A, Gill R, Thomas DM, Gilbertson J, Foster C, Marshall CJ (1991) Multiple K-ras codon 12 mutations in cholangiocarcinomas demonstrated with a sensitive polymerase chain reaction technique. Cancer Res 51(13):3497–3502

    PubMed  CAS  Google Scholar 

  23. Ahmadian A, Ehn M, Hober S (2006) Pyrosequencing: history, biochemistry and future. Clin Chim Acta 363(1–2):83–94

    Article  PubMed  CAS  Google Scholar 

  24. Poehlmann A, Kuester D, Meyer F, Lippert H, Roessner A, Schneider-Stock R (2007) K-ras mutation detection in colorectal cancer using the Pyrosequencing technique. Pathol Res Pract 203(7):489–497

    Article  PubMed  CAS  Google Scholar 

  25. Newton CR, Graham A, Heptinstall LE, Powell SJ, Summers C, Kalsheker N, Smith JC, Markham AF (1989) Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). Nucleic Acids Res 17(7):2503–2516

    Article  PubMed  CAS  Google Scholar 

  26. Clayton SJ, Scott FM, Walker J, Callaghan K, Haque K, Liloglou T, Xinarianos G, Shawcross S, Ceuppens P, Field JK et al (2000) K-ras point mutation detection in lung cancer: comparison of two approaches to somatic mutation detection using ARMS allele-specific amplification. Clin Chem 46(12):1929–1938

    PubMed  CAS  Google Scholar 

  27. Whitcombe D, Theaker J, Guy SP, Brown T, Little S (1999) Detection of PCR products using self-probing amplicons and fluorescence. Nat Biotechnol 17(8):804–807

    Article  PubMed  CAS  Google Scholar 

  28. Thelwell N, Millington S, Solinas A, Booth J, Brown T (2000) Mode of action and application of Scorpion primers to mutation detection. Nucleic Acids Res 28(19):3752–3761

    Article  PubMed  CAS  Google Scholar 

  29. Oliner K, Juan T, Suggs S, Wolf M, Sarosi I, Freeman DJ, Gyuris T, Baron W, Bakker A, Parker A et al (2010) A comparability study of 5 commercial KRAS tests. Diagn Pathol 5:23–30

    Article  PubMed  Google Scholar 

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Acknowledgments

Thank to Oncology, Pathology and Research Unit in General Yagüe Hospital in Burgos, Spain. MHV is especially thankful to CVP, IHH and AHV, for their support.

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Authors and Affiliations

  1. Research Unit, Hospital General Yagüe, Avenida del Cid 96, 09005, Burgos, Spain

    Marta Herreros-Villanueva

  2. Division of Gastroenterology and Hepatology, Department of Medicine, Mayo Clinic, Rochester, MN, USA

    Gaurav Aggarwal

Authors
  1. Marta Herreros-Villanueva
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  2. Gaurav Aggarwal
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Correspondence to Marta Herreros-Villanueva.

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Herreros-Villanueva, M., Aggarwal, G. KRAS assay selection: sensitivity and accuracy in clinical application. Mol Biol Rep 39, 2467–2470 (2012). https://doi.org/10.1007/s11033-011-0997-6

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  • DOI: https://doi.org/10.1007/s11033-011-0997-6

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