Utility of a limited panel of calretinin and Ber-EP4 immunocytochemistry on cytospin preparation of serous effusions: A cost-effective measure in resource-limited settings

Abstract

Background: Differentiation between reactive, but morphologically atypical, mesothelial cells and adenocarcinoma in effusions can be problematic. Elaborate immunohistochemical panels have been devised. Techniques like DNA analysis, flow/image cytometry, and K-ras mutation analysis are research oriented and difficult to perform in routine, especially in resource-poor centers. We evaluated the efficacy of a limited two-antibody panel comprising calretinin and Ber-EP4 on cytospin and cell block preparations, in 100 effusion samples.Materials and Methods: Fifty cases of reactive mesothelial hyperplasia and 50 cases of adenocarcinoma diagnosed by cytomorphology in ascitic/pleural fluid specimens over a 2-year period were assessed. The diagnoses were confirmed by clinical/histopathologic correlation. Cytospin smears were made in all. Cell blocks were prepared, wherever adequate fluid was available. Immunocytochemistry (ICC) for calretinin and Ber-EP4 was performed.Results: Forty-five of the reactive effusion cases (90%) were calretinin reactive and Ber-EP4 negative. Among the adenocarcinoma cases, 49 (98%) were calretinin negative but Ber-EP4 positive. Thus, both calretinin and Ber-EP4 had a high sensitivity (90% and 98%, respectively), as well as a high specificity (100% and 86%, respectively). In the 21 reactive mesothelial cases, whose cell blocks were made, results were comparable to those on cytospin. However, of the 19 adenocarcinoma cases in which cell blocks were prepared, all were Ber-EP4 immunopositive except for three, which were positive on cytospin, implying false-negative results on cell blocks.Conclusions: A limited panel of two monoclonal antibodies, calretinin and Ber-EP4, may be useful in cytology, as a “primary antibody panel”, for accurate diagnosis and patient management. Additionally, ICC can be performed easily on cytospin preparations, which gave results comparable to cell blocks in our study.

The Use of Sensitive Chemical Antibodies for Diagnosis: Detection of Low Levels of Epcam in Breast Cancer

“…This discrepancy in staining is not a phenomenon solely associated with aptamers and is observed with antibodies as well. In the case of EpCAM detection, the Ber-EP4 antibody has been shown to display false negative results in paraffin embedded cell blocks [43]. Therefore, as with antibodies, a negative result needs to be confirmed with additional aptamers to confirm the lack of expression, rather than the lack of a reaction [44].…”

Cytologic and Immunophenotypic Features of Malignant Cells in Pediatric Body Fluids

“…In general, ICC is valuable in the management of malignant effusions and is performed on cell blocks and cytospin smears [11,12]. The use of calretinin and epithelial markers has been emphasized in serous effusions and cerebrospinal fluid with little or no false-positive/-negative results [13,14,15]. Similar results were encountered in our study with the use of cytokeratins and calretinin, which gave the best performance of all antibody tests, i.e.…”

Cell blocks in cytopathology: a review of preparative methods, utility in diagnosis and role in ancillary studies

“…CBs are the preferred choice for ICC as they are comparable with surgical biopsies, can use the same control slides, provide the best milieu for morphological interpretation and are without background staining. However, in low‐resource settings, cytospin preparations have been considered as a useful alternative . Appropriate tissue triaging has become an important aspect of pathology in today's era of personalized medicine to avoid subjecting patients to unnecessary sampling.…”

“…The clinical utility of cytological specimens, including CB, for molecular testing in lung cancer has been validated recently for the personalized treatment of non‐small‐cell lung carcinoma . CBs permit the analysis of molecular alterations in lung cancer using different cytogenetic and molecular technologies . The reported figures for inadequate cellularity in the molecular analysis of lung cancer vary from as low as 6.4% to as high as 57% .…”