Evaluation of a flow cytometry-based surrogate assay (FlowSA) for the detection of SARS-CoV-2 in clinical samples

IF 1.4 Q4 INFECTIOUS DISEASES Journal of clinical virology plus Pub Date : 2025-02-01 Epub Date: 2025-01-26 DOI:10.1016/j.jcvp.2025.100204

Vinit Upasani , Marjolein Knoester , Daniele Pantano , Lili Gard , Jolanda M. Smit , Bernardina T.F. van der Gun , Adriana Tami , Izabela A. Rodenhuis-Zybert

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Abstract

Introduction

The current diagnostic methods for SARS-CoV-2 rely on quantitative RT-PCR. However, the presence of viral RNA in samples does not necessarily reflect the presence of an infectious virus. Therefore, the reliable detection of infectious SARS-CoV-2 in clinical samples is necessary to limit viral transmission.

Methods

We developed a flow cytometry-based surrogate assay (FlowSA), wherein the presence of infectious SARS-CoV-2 was detected using virus nucleocapsid-specific antibodies.

Results

We showed that FlowSA allows the detection of a wide range of viral titers of multiple SARS-CoV-2 variants. Furthermore, the assay was successfully used to detect infectious SARS-CoV-2 in nasopharyngeal swabs from SARS-CoV-2 positive individuals, including those with high Ct values. Notably, FlowSA identified the presence of infectious SARS-CoV-2 in biological specimens that scored negative for cytopathic effect (CPE) in cell culture and would otherwise be considered negative.

Conclusion

We propose that FlowSA can be adopted as an alternative to conventional CPE methods for viral diagnostics.

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基于流式细胞术的替代检测法（FlowSA）在临床样品中检测SARS-CoV-2的评价

目前SARS-CoV-2的诊断方法主要依赖于定量RT-PCR。然而，样本中病毒RNA的存在并不一定反映感染性病毒的存在。因此，在临床样本中可靠地检测传染性SARS-CoV-2是限制病毒传播的必要条件。方法我们建立了一种基于流式细胞术的替代检测方法（FlowSA），其中使用病毒核衣壳特异性抗体检测传染性SARS-CoV-2的存在。结果FlowSA可以检测多种SARS-CoV-2变体的大范围病毒滴度。此外，该方法还成功地用于检测SARS-CoV-2阳性个体（包括Ct值高的个体）鼻咽拭子中的传染性SARS-CoV-2。值得注意的是，FlowSA在细胞培养中细胞病变效应（CPE）得分为阴性的生物标本中发现了传染性SARS-CoV-2的存在，否则将被视为阴性。结论FlowSA可作为传统CPE方法的替代方法用于病毒诊断。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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