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Standardization of the immunocytochemical detection of cancer cells in BM and blood: I. establishment of objective criteria for the evaluation of immunostained cells

DOI: 10.1080/0032472031000141283
  • Micrometastasis
  • Bone Marrow
  • Immunocytochemistry
  • Morphology
  • Standardization
  • Classification
  • Carcinoma
  • Neoplasm Metastasis
  • Chemistry
  • Medicine


Background Detection of isolated tumor cells (TC) in BM from carcinoma patients can predict future relapse. Various molecular and immunocytochemical (ICC) methods have been used to detect these cells, which are present at extremely low frequencies of 10−5−10−6. The specificity and sensitivity of these techniques may vary widely. In 1996, a European ISHAGE Working Group was founded to standardize and optimize procedures used for the detection of minimal residual disease. We have attempted to develop objective criteria for the evaluation of immunocytochemi-cally identifiable cancer cells. Methods An interlaboratory ring experiment was performed, to compare the screening and detection of micrometastasis-positive events between different laboratories. The discrepant results induced us to establish a common consensus on morphological criteria applicable to the identification of immunostained micrometastatic TC. Results Bared on this consensus evaluation, we propose a classification of stained elements into three groups: (1) ‘TC's show patbognomonic signs of epithelial TC-nature, as defined by a clearly enlarged nucleus or clusters of ≥ 2 immunopositive cells. (2) ‘Probable TC's represent morphological overlap between hematopoietic cells (HC) and TC which lack pathognomonic signs of TC-nature, but do not exhibit clear morphological features of HC. These cells are considered as TC if control staining with an isotype-specific, unrelated Ab is negative. (3) ‘TC-negative’ cells are defined as false positive’ HC, skin squamous epithelial cells and artefacts. Discussion The proposed classification of immunostained events is a first step towards the development of standardized immunocytochemical assays for the detection of occult micrometastatic TC in BM or blood.

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