FFPE tissue was sectioned and stained for SDHB using rabbit polyclonal anti-SDHB (HPA002868, 1:400 dilution; Sigma-Aldrich, St Louis, MO, USA) according to the local procedures of six different centres (Dresden, Bethesda, Madrid, Florence, Nijmegen, and Rotterdam) [9 (link)]. For 23% of tumours (44 samples), a tissue microarray was constructed with three cores of 1.0 mm per sample. Local pathologists evaluated SDHB staining in one slide per tumour and gave the results in four categories: as positive, for the typical granular staining pattern; as negative, for completely negative or weak diffuse staining; as inconclusive, when both patterns were present; or as non-informative, when tissue or staining artefacts were observed. For 50 samples, the SDHB-IHC interpretations were as established previously, from combined interpretations of seven expert pathologists [20 (link)]; for these cases, results were rated as inconclusive when fewer than five pathologists agreed. All pathologists were blinded to the genetic status.
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SDHB Immunohistochemistry in Tumours
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Corresponding Organization : University Hospital Carl Gustav Carus
Other organizations : National Institutes of Health, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Spanish National Cancer Research Centre, Ludwig-Maximilians-Universität München, Erasmus MC, National Institute of Neurological Disorders and Stroke, University of Florence, Universitätsklinikum Würzburg, University of Würzburg, Städtisches Klinikum Dresden, Radboud University Nijmegen, Radboud University Medical Center, Helmholtz-Zentrum Dresden-Rossendorf, Centro de Investigación Biomédica en Red, Centre for Biomedical Network Research on Rare Diseases, Laboratoire National de Santé, Royal North Shore Hospital, University of Sydney, Tufts University, University Medical Center Utrecht, Princess Máxima Center, University of Birmingham
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Variable analysis
independent variables
- Tissue sectioning method
- Staining protocol for SDHB (antibody, dilution, and procedure)
- SDHB staining pattern (positive, negative, inconclusive, or non-informative)
- Genetic status of samples (blinded to pathologists)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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