Biochip-SER Test System for Fluorescent Immunocytochemical Analysis of Ep-CAM Antigen Expression in Effusions and Washes

The aim of the study was to evaluate the Biochip-SER test system for its ability to measure the expression of Ep-CAM antigen by fluorescent immunocytochemistry.

Materials and Methods. We conducted 64 cytological, 64 standard immunocytochemical (SICC) and 64 fluorescent immunocytochemical (FICC) tests using the Biochip-SER test system and samples from 59 patients (45 effusions and 19 peritoneal washings). The study was performed in 4 stages: cytological examination, SICC, FICC using the Biochip-SER system, and control cytological study by the Biochip-SER system. The SICC was performed using the monoclonal antibody (MAB) to the epithelial antigen Ep-CAM; FICC was performed using the MAB to the epithelial antigen Ep-CAM conjugated to the Alexa Flour 488 fluorochrome.

Results. FICC test performed using the Biochip-SER test system, showed 100% diagnostic sensitivity and 89% specificity.

Conclusion. FICC test carried out with MAB to the Ep-CAM antigen conjugated with Alexa Flour 488 using the Biochip-SER system is a reliable method for diagnosing tumors by testing effusion fluids, which allows us to recommend this system for practical use in specialized institutions and primary points of care.

1. Semenov D.A., Tseluyko S.S. Histophysiology of the pleural cavity and pleural effusion. Dal’nevostochnyy meditsinskiy zhurnal 2012; 2: 140–144.
2. Dolgov V.V., Shabalova I.P., Mironova I.I., Djangirova T.V., Korotaev A.L. Vypotnye zhidkosti. Laboratornoe issledovanie [Serous effusions. Laboratory diagnostics]. Moscow: Triada; 2006.
3. Gluzman D.F., Sklyarenko L.M., Nadgornaya V.A., Kryachok I.A. Diagnosticheskaya immunotsitokhimiya opukholey [Diagnostic immunocytochemistry of tumors]. Kiev: Morion; 2003.
4. Shidham V.B., Atkinson B.F. Cytopathologic diagnosis of serous fluids. Elsevier Saunders; 2007.
5. Savostikova M.V., Furminskaya E.Yu., Fedoseeva E.S., Kudaibergenova A.G. The fluorescent immune cytochemical analysis of exudations and washouts from serous cavities under intra-operational cytological diagnostic. Klinicheskaia laboratornaia diagnostika 2017; 62(12): 742–745.
6. Lazarev A.F., Grigoruk O.G., Bazulina L.M., Muzalevskiy P.N., Kravtsov V.Yu. Pleural mesothelioma: etiology, incidence, diagnosis, treatment and survival. Rossiyskiy onkologicheskiy zhurnal 2013; 5: 15–20.
7. Davidson B. Malignant nonhematological effusion characterization by flow cytometry. Acta Cytol 2016; 60(4): 365–371, https://doi.org/10.1159/000447687.
8. Bolgova L.S., Marinenko S.V. Modern possibilities of differential cytological diagnostics of proliferating mesothelium, mesothelioma and cancer metastases (literature review and results of own research). Klinicheskaya onkologiya 2015; 4(20).
9. Gabris S., Kern L. Two color immunostaining of pleural effusions with Ber-EP4 and CK5/6. Cytopathology 2004; 15(Suppl 2): 14.
10. Comin C.E., Saieva C., Messerini L. H-caldesmon, calretinin, estrogen receptor, and Ber-EP4: a useful combination of immunohistochemical markers for differentiating epithelioid peritoneal mesothelioma from serous papillary carcinoma of the ovary. Am J Surg Pathol 2007; 31(8): 1139–1148, https://doi.org/10.1097/pas.0b013e318033e7a8.
11. Schnell U., Cirulli V., Giepmans B.N. EpCAM: structure and function in health and disease. Biochim Biophys Acta 2013; 1828(8): 1989–2001, https://doi.org/10.1016/j.bbamem.2013.04.018.
12. Latza U., Niedobitek G., Schwarting R., Nekarda H., Stein H. Ber-EP4: new monoclonal antibody which distinguishes epithelia from mesothelial. J Clin Pathol 1990; 43(3): 213–219, https://doi.org/10.1136/jcp.43.3.213.
13. Wang B., Li D., Ou X., Yi Q., Feng Y. Diagnostic accuracy of Ber-EP4 for metastatic adenocarcinoma in serous effusions: a meta-analysis. PLoS One 2014; 9(9): e107741, https://doi.org/10.1371/journal.pone.0107741.
14. Maguire B., Whitaker D., Carrello S., Spagnolo D. Monoclonal antibody Ber-EP4: its use in the differential diagnosis of malignant mesothelioma and carcinoma in cell blocks of malignant effusions and FNA specimens. Diagn Cytopathol 1994; 10(2): 130–134, https://doi.org/10.1002/dc.2840100207.
15. Volchenko N.N., Borisova O.V. Role of epithelial antigen Ber-EP4 in the study of exudates from serous sacs. Rossiyskiy onkologicheskiy zhurnal 2012; 2: 18–22.
16. Ordóñez N.G. Value of the Ber-EP4 antibody in differentiating epithelial pleural mesothelioma from adenocarcinoma: the M.D. Anderson experience and a critical review of the literature. Am J Clin Pathol 1998; 109(1): 85–89, https://doi.org/10.1093/ajcp/109.1.85.
17. Ordóñez N.G. The Immunohistochemical diagnosis of mesothelioma. Am J Surg Pathol 2003; 27(8): 1031–1051, https://doi.org/10.1097/00000478-200308000-00001.
18. Morimoto A., Ito A., Hashimoto K., Nakano A., Nagasaka T., Yokoi T. New diagnostic technique for rapid fluorescence immunocytochemical staining of adenocarcinoma and mesothelial cells using liquid-based cytology. Acta Cytol 2014; 58(5): 461–468, https://doi.org/10.1159/000367706.
19. Zinoviev S.V., Rachkov V.V., Utkin O.V., Savostikova M.V., Furminskaya E.Yu. A biochip for multiplex analysis and a method of studying cells in the diagnosis of oncological diseases. Patent WO 2017/204674 A1. 2017.

Zinoviev S.V., Terentiev I.G., Utkin O.V., Furminskaya E.Yu., Fedoseeva E.S., Savostikova M.V. Biochip-SER Test System for Fluorescent Immunocytochemical Analysis of Ep-CAM Antigen Expression in Effusions and Washes. Sovremennye tehnologii v medicine 2018; 10(4): 34, https://doi.org/10.17691/stm2018.10.4.04