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

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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引用次数: 0

摘要

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。

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

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.