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Study of KRAS new predictive marker in a clinical laboratory

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Abstract

Background

The presence of somatic mutations in the KRAS gene has been identified as a reliable strong negative predictor for the response to targeting the epidermal growth factor receptor (EGFR), in patients with metastatic colorectal cancer and the use of anti-EGFR monoclonal antibodies such as Cetuximab and Panitumumab is now restricted to patients with no detectable KRAS mutations. Between 30 and 40 % of colorectal cancers contain a mutated KRAS oncogene. The aim of this study was to evaluate concordance between three methods to analyze KRAS mutational status in regard to clinical testing.

Methods

We analyzed KRAS mutations in codons 12 and 13 of exon 2 in one hundred formalin-fixed paraffin-embedded (FFPE) colorectal cancer samples by three different methods: Direct Sequencing and two commercial kits on allele-specific oligonucleotide hybridization (KRAS StripAssay, Vienna Lab.) and Amplification Refractory Mutation System/Scorpions (ARMS/S; TheraScreen KRAS Mutation kit DxS) based on q-PCR.

Results

We have found similar frequencies of KRAS mutations by TheraScreen and Strip-Assay (44 and 48 %), with a κ value of 0.90, indicating almost perfect agreement between methods. The frequency by direct sequencing was much lower (26 %) and the κ values were 0.67 (compared to TheraScreen) and 0.57 (compared to Strip-Assay) indicating low sensitivity.

Conclusions

On analyzing KRAS mutation in FFPE tumor samples, direct sequencing sensitivity is too low to be used in a clinical setting. Choosing between ARMS/S; TheraScreen KRAS Mutation kit DxS and KRAS StripAssay, Vienna Lab, will depend on laboratory facilities and expertise.

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Acknowledgments

We wish to thank Jesus Sanchez for his statistical support. The present study was supported by research grants RTICC 06/0020/0021, FIS 09/00838 (Instituto de Salud Carlos III) and “Proyecto Determina KRAS” supported by MERCK.

Conflict of interest

No competing interests are declared.

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Authors and Affiliations

  1. Laboratorio de Oncología Molecular, Planta Baja Sur, Hospital Clínico San Carlos, C/Martín Lagos s/n, Madrid, 28040, Spain

    Inmaculada Bando, Lourdes Cillero, Patricia Llovet, Paula Pescador, Miguel de la Hoya & Trinidad Caldés

  2. Anatomical Pathology, Hospital Clínico San Carlos, Madrid, Spain

    Julián Sanz-Ortega & Milagros Ferrer

  3. Medical Oncology, Hospital Clínico San Carlos, Madrid, Spain

    Javier Sastre & Eduardo Díaz-Rubio García

Authors
  1. Inmaculada Bando
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  2. Lourdes Cillero
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  3. Julián Sanz-Ortega
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  4. Patricia Llovet
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  5. Paula Pescador
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  6. Milagros Ferrer
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  7. Miguel de la Hoya
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  8. Javier Sastre
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  9. Eduardo Díaz-Rubio García
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  10. Trinidad Caldés
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Corresponding author

Correspondence to Trinidad Caldés.

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Bando, I., Cillero, L., Sanz-Ortega, J. et al. Study of KRAS new predictive marker in a clinical laboratory. Clin Transl Oncol 14, 937–942 (2012). https://doi.org/10.1007/s12094-012-0886-z

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  • DOI: https://doi.org/10.1007/s12094-012-0886-z

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