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Molecular Biology Reports

, Volume 39, Issue 3, pp 2467–2470 | Cite as

KRAS assay selection: sensitivity and accuracy in clinical application

  • Marta Herreros-Villanueva
  • Gaurav Aggarwal
Article

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.

Keywords

KRAS Mutations Sensitivity Accuracy Specificity Mutation analysis Colorectal cancer EGFR inhibitors 

Notes

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.

Conflict of interest

None.

References

  1. 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):2040–2048PubMedCrossRefGoogle Scholar
  2. 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):92–98PubMedCrossRefGoogle Scholar
  3. 3.
    Malumbres M, Barbacid M (2003) RAS oncogenes: the first 30 years. Nat Rev 3(6):459–465CrossRefGoogle Scholar
  4. 4.
    Jancik S, Drabek J, Radzioch D, Hajduch M (2010) Clinical relevance of KRAS in human cancers. J Biomed Biotechnol 2010:150960PubMedCrossRefGoogle Scholar
  5. 5.
    Schubbert S, Shannon K, Bollag G (2007) Hyperactive Ras in developmental disorders and cancer. Nat Rev 7(4):295–308CrossRefGoogle Scholar
  6. 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):1626–1634PubMedCrossRefGoogle Scholar
  7. 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):1315–1320PubMedCrossRefGoogle Scholar
  8. 8.
    Venook AP (2005) Epidermal growth factor receptor-targeted treatment for advanced colorectal carcinoma. Cancer 103(12):2435–2446PubMedCrossRefGoogle Scholar
  9. 9.
    Kohne CH (2010) How to integrate molecular targeted agents in the continuum of care. Ann Oncol 21(Suppl 7):134–139Google Scholar
  10. 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):5931–5937PubMedCrossRefGoogle Scholar
  11. 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):1254–1264PubMedCrossRefGoogle Scholar
  12. 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):413–421PubMedCrossRefGoogle Scholar
  13. 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):254–259PubMedCrossRefGoogle Scholar
  14. 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 8:142–156PubMedCrossRefGoogle Scholar
  15. 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):e53PubMedCrossRefGoogle Scholar
  16. 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 1022:250–256PubMedCrossRefGoogle Scholar
  17. 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):508–513PubMedCrossRefGoogle Scholar
  18. 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):197–205PubMedCrossRefGoogle Scholar
  19. 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):162–170PubMedGoogle Scholar
  20. 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):8093–8099PubMedCrossRefGoogle Scholar
  21. 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):923–928PubMedGoogle Scholar
  22. 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):3497–3502PubMedGoogle Scholar
  23. 23.
    Ahmadian A, Ehn M, Hober S (2006) Pyrosequencing: history, biochemistry and future. Clin Chim Acta 363(1–2):83–94PubMedCrossRefGoogle Scholar
  24. 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):489–497PubMedCrossRefGoogle Scholar
  25. 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):2503–2516PubMedCrossRefGoogle Scholar
  26. 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):1929–1938PubMedGoogle Scholar
  27. 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):804–807PubMedCrossRefGoogle Scholar
  28. 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):3752–3761PubMedCrossRefGoogle Scholar
  29. 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 5:23–30PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.Research UnitHospital General YagüeBurgosSpain
  2. 2.Division of Gastroenterology and Hepatology, Department of MedicineMayo ClinicRochesterUSA

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