How to cite this article: Kentrou NA, Tsagarakis NJ, Tzanetou K, Damala M, Papadimitriou KA, Skoumi D, Stratigaki A, Anagnostopoulos NI, Malamou-Lada E, Athanassiadou P, Paterakis G. An improved flow cytometric assay for detection and discrimination between malignant cells and atypical mesothelial cells, in serous cavity effusions. Cytometry Part B 2011; 80B: 324–334.
An improved flow cytometric assay for detection and discrimination between malignant cells and atypical mesothelial cells, in serous cavity effusions
Article first published online: 21 JUN 2011
Copyright © 2011 International Clinical Cytometry Society
Cytometry Part B: Clinical Cytometry
Volume 80B, Issue 5, pages 324–334, September 2011
How to Cite
Kentrou, N. A., Tsagarakis, N. J., Tzanetou, K., Damala, M., Papadimitriou, K. A., Skoumi, D., Stratigaki, A., Anagnostopoulos, N. I., Malamou-Lada, E., Athanassiadou, P. and Paterakis, G. (2011), An improved flow cytometric assay for detection and discrimination between malignant cells and atypical mesothelial cells, in serous cavity effusions. Cytometry, 80B: 324–334. doi: 10.1002/cyto.b.20608
- Issue published online: 17 AUG 2011
- Article first published online: 21 JUN 2011
- Accepted manuscript online: 25 MAY 2011 02:30PM EST
- Manuscript Accepted: 19 MAY 2011
- Manuscript Revised: 13 MAY 2011
- Manuscript Received: 19 FEB 2011
- flow cytometry;
- malignant effusions;
- atypical mesothelial cells
The aim of this study was to evaluate a flow cytometric assay for the detection of malignant effusions.
During the last 4-year period, 125 effusions suspicious for malignancy were prospectively analyzed by flow cytometry and conventional cytology. A three-step flow cytometric assay was performed, beginning with an initial informative panel of two protocols, containing SYTO-16, 7-AAD, CD71-PE, CD45-ECD, and CD66abce-FITC, CD64-PE, CD45-ECD, CD16-PECy5, CD14-PECy7, respectively. This was followed by a basic immunophenotypic panel of seven three-color combinations, containing in the first position, EMA, Ber-EP4, CD66abce, CD56, and intracellular desmin-33, combined with CD71-PE and CD45-PeCy5 in each tube. Finally, a cytokeratin-FITC/propidium iodide DNA panel was conducted, for the detection of aneuploidy in cytokeratin positive cells.
The sensitivity and specificity of flow cytometry were 85.1 and 97.8%, and of cytology 93.2 and 95.6%, respectively. A significant association was observed between the results of the two techniques (P < 0.001). Among eight atypical cases detected by cytology, five had been precisely characterized as malignant by flow cytometry. EMA and Ber-EP4 proved the most sensitive markers for malignancy diagnosis, while the detection of desmin-33 negative/cytokeratin positive cells had the simultaneous highest positive and negative predictive values. CD66abce was very specific, although nonsensitive, while DNA ploidy analysis was nonspecific, as hyperploidy was observed in reactive mesothelial cells.
A flow cytometric assay of high sensitivity and specificity is proposed for the routine identification of carcinoma cells in effusions and their distinction from atypical mesothelial cells, as an ancillary to conventional cytology. © 2011 International Clinical Cytometry Society