title: Performance of 4 Immunohistochemical Phosphohistone H3 Antibodies for Marking Mitotic Figures in Breast Cancer.
abstract: BACKGROUND
  Phosphohistone H3 (PHH3) has been suggested to facilitate and improve mitotic activity assessment in breast cancer and other tumor entities, but the reliability of respective immunohistochemical antibodies has not yet been compared for routine purposes. Our aim was to test the performance of 4 different PHH3 antibodies on a series of highly proliferating breast cancers with good preservation of morphology.
  
  
  METHODS
  Four commercially available PHH3 antibodies were tested on 9 grade 3 invasive breast cancers processed in the same batch. We analyzed the number of antibody stained and nonstained mitotic figures as well as the total of cells observed in 10 high power fields per tumor to calculate sensitivity, specificity, and accuracy of the respective antibodies for staining mitotic figures, taking morphologically defined mitotic figures as gold standard.
  
  
  RESULTS
  Sensitivity, specificity, and accuracy of the respective PHH3 antibodies for staining mitotic figures were 54.51%, 99.98%, and 98.79% for Cell Marque, 87.48%, 67.62%, and 67.47% for Epitomics, 98.62%, 99.73%, and 99.49% for Merck 06-570, and 99.74%, 99.52%, and 99.51% for Merck 09-797, respectively. Sensitivity was lowest for telophase. In statistical analysis, the Cell Marque antibody demonstrated significantly lower sensitivity and Epitomics substantially lower sensitivity and specificity than Merck 06-570 and Merck 09-797 antibodies (P<0.0001, respectively).
  
  
  CONCLUSIONS
  Performance and reliability varied significantly between the 4 tested antibodies. For faster identification of mitotic hot spots and as potential marker in digital image analysis, the Merck antibodies seem to be most suitable.
authors.affiliation: Department of Pathology, Dietrich Bonhoeffer Medical Centre, Neubrandenburg, Germany.|Department of Pathology, University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Cornelia M
authors.initials: CM
authors.lastname: Focke
authors.affiliation: Department of Pathology, Dietrich Bonhoeffer Medical Centre, Neubrandenburg, Germany.
authors.forename: Kai
authors.initials: K
authors.lastname: Finsterbusch
authors.affiliation: Department of Pathology, Dietrich Bonhoeffer Medical Centre, Neubrandenburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Decker
authors.affiliation: Department of Pathology, University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: van Diest
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000473:pathology; D005260:Female; D006657:Histones / Q000737:chemistry*; D006801:Humans; D007150:Immunohistochemistry / Q000592:standards*; D057230:Limit of Detection; D008938:Mitosis; D015203:Reproducibility of Results; D013194:Staining and Labeling / Q000592:standards*
chemical_list: D000911:Antibodies, Monoclonal; D006657:Histones