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The reliability of immunohistochemistry as a prescreening method for the diagnosis of hereditary nonpolyposis colorectal cancer (HNPCC) – Results of an international collaborative study

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Abstract

Hereditary nonpolyposis colorectal cancer syndrome (HNPCC) is an autosomal dominant condition accounting for 2–5% of all colorectal carcinomas as well as a small subset of endometrial, upper urinary tract and other gastrointestinal cancers. An assay to detect the underlying defect in HNPCC, inactivation of a DNA mismatch repair enzyme, would be useful in identifying HNPCC probands. Monoclonal antibodies against hMLH1 and hMSH2, two DNA mismatch repair proteins which account for most HNPCC cancers, are commercially available. This study sought to investigate the potential utility of these antibodies in determining the expression status of these proteins in paraffin-embedded formalin-fixed tissue and to identify key technical protocol components associated with successful staining. A set of 20 colorectal carcinoma cases of known hMLH1 and hMSH2 mutation and expression status underwent immunoperoxidase staining at multiple institutions, each of which used their own technical protocol. Staining for hMSH2 was successful in most laboratories while staining for hMLH1 proved problematic in multiple labs. However, a significant minority of laboratories demonstrated excellent results including high discriminatory power with both monoclonal antibodies. These laboratories appropriately identified hMLH1 or hMSH2 inactivation with high sensitivity and specificity. The key protocol point associated with successful staining was an antigen retrieval step involving heat treatment and either EDTA or citrate buffer. This study demonstrates the potential utility of immunohistochemistry in detecting HNPCC probands and identifies key technical components for successful staining.

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Authors and Affiliations

  1. Institute of Pathology, Heinrich Heine University, Düsseldorf, Germany

    Wolfram Müller

  2. Department of Pathology, Mayo Clinic, Rochester, Minnesota, 55905, USA

    Lawrence J. Burgart

  3. Department of Surgery, Heinrich Heine University, Düsseldorf, Germany

    Ruth Krause-Paulus & Gabriela Möslein

  4. Department of Molecular Genetics, Mayo Clinic, Rochester, Minnesota, USA

    Stephen N. Thibodeau

  5. Northern Genetics Center, University of Newcastle, Newcastle upon Tyne, UK

    M. Almeida

  6. Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, USA

    T. Brocker Edmonston

  7. University of California, San Diego, California, USA

    C. R. Boland

  8. Department of Surgery, Division of Molecular Diagnostics & Therapy, University of Heidelberg, Germany

    C. Sutter

  9. Department of Pathology, Graduate Medical School, University of Queensland, Australia

    J. R. Jass

  10. Department of Clinical Genetics, Karolinska Hospital, Stockholm, Sweden

    A. Lindblom

  11. Hereditary Cancer Center, Pomerian Medical University, Szczecin, Poland

    J. Lubinski

  12. Department of Clinical Genetics, Royal Free Hospital, London, UK

    K. MacDermot

  13. Department of Pathology, The Medical School, Edgbaston, Birmingham, B15 2TT, UK

    D. S. A. Sanders

  14. Department of Pathology, University of Leiden, The Netherlands

    H. Morreau

  15. Department of Surgery, University of Göttingen, Germany

    A. Müller

  16. Division of Oncology, Ospedale Civile Maggiore, Verona, Italy

    C. Oliani

  17. Department of Clinical Biochemistry, Aarhus University, Denmark

    T. Orntoft

  18. Department of Medicine III, University Modena, Modena, Italy

    M. Ponz De Leon

  19. Laboratory of Molecular Toxicology U490, University René Descartes, Paris V, France

    C. Rosty

  20. Department of Surgery Oncology, Roswell Park Cancer Inst., New York, USA

    M. Rodriguez-Bigas

  21. Department of Pathology, Städt. Klinikum Kassel, Kassel, Germany

    J. Rüschoff

  22. Division of Tissue Pathology, Institute of Medical + Veterinary Science, Rundle Mall, Adelaide, Australia

    A. Ruszkiewicz

  23. Department of Pathology, Institute Gustav Roussy, Villejuif Cedes, France

    J. Sabourin

  24. Department of Medical Genetics and Pathology, University of Helsinki, Helsinki, Finland

    R. Salovaara

Authors
  1. Wolfram Müller
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  2. Lawrence J. Burgart
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  3. Ruth Krause-Paulus
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  4. Stephen N. Thibodeau
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  5. M. Almeida
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  6. T. Brocker Edmonston
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  7. C. R. Boland
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  8. C. Sutter
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  9. J. R. Jass
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  10. A. Lindblom
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  11. J. Lubinski
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  12. K. MacDermot
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  13. D. S. A. Sanders
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  14. H. Morreau
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  15. A. Müller
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  16. C. Oliani
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  17. T. Orntoft
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  18. M. Ponz De Leon
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  19. C. Rosty
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  20. M. Rodriguez-Bigas
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  21. J. Rüschoff
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  22. A. Ruszkiewicz
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  23. J. Sabourin
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  24. R. Salovaara
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  25. Gabriela Möslein
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Müller, W., Burgart, L.J., Krause-Paulus, R. et al. The reliability of immunohistochemistry as a prescreening method for the diagnosis of hereditary nonpolyposis colorectal cancer (HNPCC) – Results of an international collaborative study. Familial Cancer 1, 87–93 (2001). https://doi.org/10.1023/A:1013840907881

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  • DOI: https://doi.org/10.1023/A:1013840907881

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