Simple and rapid flow cytometry assay for the detection of malignant epithelial cells in body fluids

Author:

Ramalingam Thulasi Raman1ORCID,Mani Sandeep2,Narla Swetha2,Lakshmanan Archana2,Muthu Anurekha1,Nk Harsha Rasheed1,Mohanraj Selvaraj1,Ramachandran Ellapan Dharma1,Subramanyam Annapurneswari2,Vaidhyanathan Lakshman2

Affiliation:

1. Department of Hematology Apollo Cancer Centre Chennai India

2. Department of Histopathology Apollo Cancer Centre Chennai India

Abstract

AbstractIntroductionMorphology is routinely used for detecting malignant cells in body fluids, but it has limitations. Recently, flow cytometry (FCM) is used as an effective tool for studying non‐haematological malignancies. The main objective of this study is to standardize a simple and rapid FCM test for the detection of malignant epithelial cells in body fluids.Materials and MethodsBody fluids that had been processed for cytology/cytology and FCM were enrolled in this prospective study. We developed a fluorescent‐labelled, monoclonal antibody panel composed of cell surface markers for this FCM assay. We compared the results of cytology/cell block and FCM.ResultsA total of 121 fluid samples were studied. Comparing the diagnostic performance of cytology/cell block and FCM, 52 (43%) cases were positive and 60 (49.5%) cases were negative for carcinoma cells by both techniques. Nine cases showed discordant results between the two techniques. Six cases were cytology+ but FCM‐ and three cases were FCM+ cytology‐. Clustered Epithelial Cell Adhesion Molecule (EpCAM)‐positive events with high scatter properties were definitive for positive diagnosis by FCM. We studied PD‐L1 expression in 13 cases by FCM. Six cases were reported as false negative by this FCM assay due to hypocellularity and lack of EpCAM expression in malignant cells.ConclusionsThis FCM assay is simple, easier and cost‐effective yielding sensitive results with no inter‐observer variability. FCM would become a valuable tool to complement routine diagnostic cytology and reduces misdiagnosis.

Publisher

Wiley

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