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

IF 1.2 4区医学 Q4 CELL BIOLOGY Cytopathology Pub Date : 2024-06-04 DOI:10.1111/cyt.13408

Thulasi Raman Ramalingam, Sandeep Mani, Swetha Narla, Archana Lakshmanan, Anurekha Muthu, Harsha Rasheed Nk, Selvaraj Mohanraj, Ellapan Dharma Ramachandran, Annapurneswari Subramanyam, Lakshman Vaidhyanathan

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Abstract

Introduction: Morphology 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 methods: Body 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.

Results: A 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.

Conclusions: This 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.

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用于检测体液中恶性上皮细胞的简单快速的流式细胞术检测方法。

简介形态学是检测体液中恶性细胞的常规方法，但有其局限性。最近，流式细胞术（FCM）被用作研究非血液学恶性肿瘤的有效工具。本研究的主要目的是将用于检测体液中恶性上皮细胞的简单快速的 FCM 检测标准化：本前瞻性研究选取了经过细胞学/细胞学和 FCM 处理的体液。我们开发了一种荧光标记的单克隆抗体面板，由细胞表面标记物组成，用于 FCM 检测。我们比较了细胞学/细胞阻断和 FCM 的结果：共研究了 121 份液体样本。比较细胞学/细胞块和 FCM 的诊断性能，两种技术的癌细胞检测结果均为阳性的病例有 52 例（43%），阴性的病例有 60 例（49.5%）。有 9 例病例两种技术的结果不一致。6 例细胞学+ 但 FCM-，3 例 FCM+ 细胞学-。上皮细胞粘附分子（EpCAM）阳性的聚集事件具有高散射特性，可确定 FCM 诊断为阳性。我们通过 FCM 研究了 13 个病例的 PD-L1 表达。有 6 例病例由于细胞功能减退和恶性细胞中缺乏 EpCAM 表达而被 FCM 检测为假阴性：结论：这种 FCM 检测方法简单、方便、成本效益高，结果灵敏，观察者之间无差异。FCM 将成为补充常规细胞学诊断的重要工具，并能减少误诊。

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来源期刊

Cytopathology 生物-病理学

CiteScore

2.30

自引率

15.40%

发文量

107

审稿时长

6-12 weeks

期刊介绍： The aim of Cytopathology is to publish articles relating to those aspects of cytology which will increase our knowledge and understanding of the aetiology, diagnosis and management of human disease. It contains original articles and critical reviews on all aspects of clinical cytology in its broadest sense, including: gynaecological and non-gynaecological cytology; fine needle aspiration and screening strategy. Cytopathology welcomes papers and articles on: ultrastructural, histochemical and immunocytochemical studies of the cell; quantitative cytology and DNA hybridization as applied to cytological material.