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 limi...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-02-01
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| Series: | Journal of Clinical Virology Plus |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667038025000031 |
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| author | Vinit Upasani Marjolein Knoester Daniele Pantano Lili Gard Jolanda M. Smit Bernardina T.F. van der Gun Adriana Tami Izabela A. Rodenhuis-Zybert |
| author_facet | Vinit Upasani Marjolein Knoester Daniele Pantano Lili Gard Jolanda M. Smit Bernardina T.F. van der Gun Adriana Tami Izabela A. Rodenhuis-Zybert |
| author_sort | Vinit Upasani |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-ab87a4c530c149698a2f3d4010d048b4 |
| institution | Kabale University |
| issn | 2667-0380 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Clinical Virology Plus |
| spelling | doaj-art-ab87a4c530c149698a2f3d4010d048b42025-02-08T05:01:33ZengElsevierJournal of Clinical Virology Plus2667-03802025-02-0151100204Evaluation of a flow cytometry-based surrogate assay (FlowSA) for the detection of SARS-CoV-2 in clinical samplesVinit Upasani0Marjolein Knoester1Daniele Pantano2Lili Gard3Jolanda M. Smit4Bernardina T.F. van der Gun5Adriana Tami6Izabela A. Rodenhuis-Zybert7University of Groningen, University Medical Center Groningen, Department of Medical Microbiology and Infection Prevention, Groningen, The NetherlandsUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology and Infection Prevention, Groningen, The NetherlandsUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology and Infection Prevention, Groningen, The Netherlands; University Hospital Münster, Institute of Hygiene, Münster, GermanyUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology and Infection Prevention, Groningen, The NetherlandsUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology and Infection Prevention, Groningen, The NetherlandsUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology and Infection Prevention, Groningen, The NetherlandsUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology and Infection Prevention, Groningen, The Netherlands; Corresponding authors.University of Groningen, University Medical Center Groningen, Department of Medical Microbiology and Infection Prevention, Groningen, The Netherlands; Corresponding authors.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.http://www.sciencedirect.com/science/article/pii/S2667038025000031SARS-CoV-2Flow cytometry-based surrogate assayNucleocapsidRT-PCRCytopathic effect |
| spellingShingle | Vinit Upasani Marjolein Knoester Daniele Pantano Lili Gard Jolanda M. Smit Bernardina T.F. van der Gun Adriana Tami Izabela A. Rodenhuis-Zybert Evaluation of a flow cytometry-based surrogate assay (FlowSA) for the detection of SARS-CoV-2 in clinical samples Journal of Clinical Virology Plus SARS-CoV-2 Flow cytometry-based surrogate assay Nucleocapsid RT-PCR Cytopathic effect |
| title | Evaluation of a flow cytometry-based surrogate assay (FlowSA) for the detection of SARS-CoV-2 in clinical samples |
| title_full | Evaluation of a flow cytometry-based surrogate assay (FlowSA) for the detection of SARS-CoV-2 in clinical samples |
| title_fullStr | Evaluation of a flow cytometry-based surrogate assay (FlowSA) for the detection of SARS-CoV-2 in clinical samples |
| title_full_unstemmed | Evaluation of a flow cytometry-based surrogate assay (FlowSA) for the detection of SARS-CoV-2 in clinical samples |
| title_short | Evaluation of a flow cytometry-based surrogate assay (FlowSA) for the detection of SARS-CoV-2 in clinical samples |
| title_sort | evaluation of a flow cytometry based surrogate assay flowsa for the detection of sars cov 2 in clinical samples |
| topic | SARS-CoV-2 Flow cytometry-based surrogate assay Nucleocapsid RT-PCR Cytopathic effect |
| url | http://www.sciencedirect.com/science/article/pii/S2667038025000031 |
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