Clinical Research Papers:

RAS testing in metastatic colorectal cancer: excellent reproducibility amongst 17 Dutch pathology centers

Annemarie Boleij, Bastiaan B.J. Tops, Paul D.M. Rombout, Elizabeth M. Dequeker, Marjolijn J.L. Ligtenberg, J. Han van Krieken _ and Dutch RAS EQA Initiative

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Oncotarget. 2015; 6:15681-15689. https://doi.org/10.18632/oncotarget.3804

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Annemarie Boleij1, Bastiaan B.J. Tops2, Paul D.M. Rombout1, Elizabeth M. Dequeker3, Marjolijn J.L. Ligtenberg1,2, J. Han van Krieken1 and Dutch RAS EQA Initiative4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20

1 Department of Pathology, Radboud University Medical Center, Nijmegen, The Netherlands

2 Department of Human Genetics, Radboud University Medical Center, Nijmegen, The Netherlands

3 Department of Public Health and Primary Care, Biomedical Quality Assurance Research Unit, KU Leuven - University of Leuven, Leuven, Belgium

4 C.J.M. van Noesel, Academic Medical Center (AMC), Department of Pathology, Amsterdam

5 C.C. Scheidel-Jacobse (Technical specialist), J.A. Kummer (Pathologist/KMBP), P. Roepman (KMBP in training), St. Antonius Ziekenhuis, Department of Pathology, Nieuwegein

6 C.F.M. Prinsen (KMBP), S.H.M. van den Berg-van Erp (Pathologist), Canisius Wilhelmina Ziekenhuis (CWZ), Department of Pathology, Nijmegen

7 J.M.H.H. van Gorp, Diakonessenhuis, Laboratory for Pathology, Utrecht

8 P.M. Nederlof, Dutch Cancer Institute (NKI), Amsterdam.

9 E. Caspers, St. Elisabeth Ziekenhuis, Department of Molecular Pathology, Tilburg

10 W.N.M. Dinjens, E.C.W. Beerens, Erasmus MC, Department of Pathology, Molecular Diagnostics, Rotterdam

11 N.A. ‘t Hart, Isala, Department of Pathology, Zwolle

12 A.J.C. van den Brule, Jeroen Bosch Ziekenhuis, Molecular Diagnostics, ‘s-Hertogenbosch

13 R. van der Geize (KMBP), S.A. Riemersma (Pathologist), Laboratory for Pathology Oost-Nederland (LABPON), Hengelo

14 T. van Wezel (KMBP), H. Morreau (Pathologist), R. van Eijk (Technical specialist), Leiden University Medical Center (LUMC), Department of Pathology, Leiden

15 J.W.M. Jeuken, Laboratory for Pathology and Medical Microbiology (PAMM), Eindhoven

16 A. Dirkx, Pathan B.V., Molecular Diagnostics, Rotterdam

17 M. Klomp, Rijnstate Ziekenhuis, Department of Pathology, Arnhem

18 W.T.M van Blokland, University Medical Center (UMC) Utrecht, Molecular Pathology, Utrecht

19 A. ter Elst (Technical specialist/KMBP in training), E. Schuuring (KMBP), A. Diepstra (Pathologist), University Medical Center Groningen (UMCG), Department of Pathology, Groningen

20 D.A.M. Heideman (KMBP), N.C.T. van Grieken (Pathologist), D. Sie (Technical specialist), VU-University Medical Center (VUMC), Department of Pathology, Amsterdam

Correspondence to:

J.Han van Krieken, email:

Keywords: RAS, colorectal cancer, metastasis, quality control, next generation sequencing

Received: February 09, 2015 Accepted: March 18, 2015 Published: April 12, 2015


In 2013 the European Medicine Agency (EMA) restricted the indication for anti-EGFR targeted therapy to metastatic colorectal cancer (mCRC) with a wild-type RAS gene, increasing the need for reliable RAS mutation testing. We evaluated the completeness and reproducibility of RAS-testing in the Netherlands.

From 17 laboratories, tumor DNA of the first 10 CRC cases tested in 2014 in routine clinical practice was re-tested by a reference laboratory using a custom next generation sequencing panel. In total, 171 CRC cases were re-evaluated for hotspot mutations in KRAS, NRAS and BRAF.

Most laboratories had introduced complete RAS-testing (65%) and BRAF-testing (71%) by January 2014. The most employed method for all hotspot regions was Sanger sequencing (range 35.7 – 49.2%). The reference laboratory detected all mutations that had been found in the participating laboratories (n = 92), plus 10 additional mutations. This concerned three RAS and seven BRAF mutations that were missed due to incomplete testing of the participating laboratory. Overall, the concordance of tests performed by both the reference and participating laboratory was 100% (163/163; κ-static 1.0) for RAS and 100% (144/144; κ-static 1.0) for BRAF.

Our study shows that RAS and BRAF mutations can be reproducibly assessed using a variety of testing methods.

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