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EGFR and KRAS quality assurance schemes in pathology: generating normative data for molecular predictive marker analysis in targeted therapy
  1. Erik Thunnissen1,
  2. Judith V M G Bovée2,
  3. Hans Bruinsma3,
  4. Adriaan J C van den Brule4,
  5. Winand Dinjens5,
  6. Daniëlle A M Heideman1,
  7. Els Meulemans6,
  8. Petra Nederlof7,
  9. Carel van Noesel8,
  10. Clemens F M Prinsen9,
  11. Karen Scheidel10,
  12. Peter M van de Ven11,
  13. Roel de Weger12,
  14. Ed Schuuring13,
  15. Marjolijn Ligtenberg14,15
  1. 1Department of Pathology, Vrije Universteit Medical Centre, Amsterdam, The Netherlands
  2. 2Department of Pathology, Leiden University Medical Centre, Leiden, The Netherlands
  3. 3Department of Pathology, Sint Elisabeth Ziekenhuis, Tilburg, The Netherlands
  4. 4Department of Pathology, PAMM Laboratory, Eindhoven, The Netherlands
  5. 5Department of Pathology, Erasmus University Medical Centre, Rotterdam, The Netherlands
  6. 6Department of Pathology, Academic Medical Centre, Maastricht, The Netherlands
  7. 7Department of Pathology, Nederlands Kanker Instituut, Amsterdam, The Netherlands
  8. 8University of Amsterdam, Department of Pathology, Academic Medical Centre, Amsterdam, The Netherlands
  9. 9Department of Pathology, Canisius Wilhelmina Ziekenhuis, Nijmegen, The Netherlands
  10. 10Department of Pathology, Antonius Ziekenhuis, Nieuwegein, The Netherlands
  11. 11Department of Epidemiology and Biostatistics, Vrije Universiteit Medical Centre, Amsterdam, The Netherlands
  12. 12Department of Pathology, University Medical Centre, Utrecht, The Netherlands
  13. 13Department of Pathology, University Medical Centre Groningen, Groningen, The Netherlands
  14. 14Department of Pathology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
  15. 15Department of Human Genetics, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
  1. Correspondence to Dr Erik Thunnissen, Department of Pathology, VU Medical Center, De Boelelaan 1117, 1081 HV Amsterdam 1081HV, The Netherlands; e.thunnissen{at}


Introduction The aim of this study was to compare the reproducibility of epidermal growth factor receptor (EGFR) immunohistochemistry (IHC), EGFR gene amplification analysis, and EGFR and KRAS mutation analysis among different laboratories performing routine diagnostic analyses in pathology in The Netherlands, and to generate normative data.

Methods In 2008, IHC, in-situ hybridisation (ISH) for EGFR, and mutation analysis for EGFR and KRAS were tested. Tissue microarray sections were distributed for IHC and ISH, and tissue sections and isolated DNA with known mutations were distributed for mutation analysis. In 2009, ISH and mutation analysis were evaluated. False-negative and false-positive results were defined as different from the consensus, and sensitivity and specificity were estimated.

Results In 2008, eight laboratories participated in the IHC ring study. In only 4/17 cases (23%) a consensus score of ≥75% was reached, indicating that this analysis was not sufficiently reliable to be applied in clinical practice. For EGFR ISH, and EGFR and KRAS mutation analysis, an interpretable result (success rate) was obtained in ≥97% of the cases, with mean sensitivity ≥96% and specificity ≥95%. For small sample proficiency testing, a norm was established defining outlier laboratories with unsatisfactory performance.

Conclusions The result of EGFR IHC is not a suitable criterion for reliably selecting patients for anti-EGFR treatment. In contrast, molecular diagnostic methods for EGFR and KRAS mutation detection and EGFR ISH may be reliably performed with high accuracy, allowing treatment decisions for lung cancer.

  • breast cancer
  • cancer genetics
  • cancer research
  • comparative genomic hybridisation
  • diagnostics
  • EGFR mutation
  • familial cancers
  • gene amplification
  • genetics
  • image analysis
  • immunohistochemistry
  • in-situ hybridisation
  • lung cancer
  • molecular pathology
  • oncology
  • proficiency testing
  • sputum

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  • Competing interests None.

  • Ethics approval Ethics approval was obtained.

  • Provenance and peer review Not commissioned; externally peer reviewed.