Development of a high-sensitivity vertical flow immunoassay for the detection of Rift Valley fever virus
ABSTRACT Rift Valley fever (RVF) is a vector-borne, zoonotic infectious disease with a proven history of morbidity and mortality in both humans and animals. Rift Valley fever virus (RVFV) is categorized as a high-priority biothreat agent by the Centers for Disease Control and Prevention and poses a...
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American Society for Microbiology
2025-07-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.00341-25 |
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| author | Alexander J. Summers Haydon J. Hill Jasmine P. Devadhasan Jian Gu Vanessa Berner Sujata G. Pandit Marcellene A. Gates-Hollingsworth Kathryn J. Pflughoeft Douglas C. Montgomery Supriya Atta Tuan Vo-Dinh David P. AuCoin Frederic Zenhausern |
| author_facet | Alexander J. Summers Haydon J. Hill Jasmine P. Devadhasan Jian Gu Vanessa Berner Sujata G. Pandit Marcellene A. Gates-Hollingsworth Kathryn J. Pflughoeft Douglas C. Montgomery Supriya Atta Tuan Vo-Dinh David P. AuCoin Frederic Zenhausern |
| author_sort | Alexander J. Summers |
| collection | DOAJ |
| description | ABSTRACT Rift Valley fever (RVF) is a vector-borne, zoonotic infectious disease with a proven history of morbidity and mortality in both humans and animals. Rift Valley fever virus (RVFV) is categorized as a high-priority biothreat agent by the Centers for Disease Control and Prevention and poses a serious national threat due to its ease of dissemination and potential for social disruption. RVF often presents as a febrile disease without specific symptoms, making early-stage detection particularly challenging. As such, it is critical that rapid, sensitive, and specific diagnostics are available for the detection of RVFV. While lateral flow immunoassays (LFIs) have been developed and validated for point-of-care (POC) diagnostics, vertical flow immunoassays (VFIs) provide enhanced analytical sensitivity and are equally suitable for POC use. In this study, we developed a VFI system for the detection of RVFV, achieving a limit of detection of 0.78 ng/mL, which is a 2.5-fold increase in analytical sensitivity compared to an LFI prototype. Furthermore, minimal cross-reactivity was demonstrated when performing the assay with target analytes of other high-priority biothreats and one other common viral nucleoprotein. This high-sensitivity VFI has the potential to prove useful for the detection of RVFV and other high-priority biothreat agents at the POC.IMPORTANCEIn this study, we have developed a rapid, sensitive vertical flow immunoassay (VFI) for the detection of Rift Valley fever virus (RVFV) in spiked human serum. The prototype diagnostic described in this research was shown to be more sensitive than traditional methods, such as lateral flow dipstick tests. Moreover, the VFI is readily deployable at the point of care in resource-limited settings. The ability of the described diagnostic to accurately and rapidly detect RVFV in samples could expedite the delivery of life-saving care and thus improve patient outcomes. |
| format | Article |
| id | doaj-art-dbe75a20129240e3a4d73983c07ac8a1 |
| institution | OA Journals |
| issn | 2165-0497 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj-art-dbe75a20129240e3a4d73983c07ac8a12025-08-20T02:38:10ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-07-0113710.1128/spectrum.00341-25Development of a high-sensitivity vertical flow immunoassay for the detection of Rift Valley fever virusAlexander J. Summers0Haydon J. Hill1Jasmine P. Devadhasan2Jian Gu3Vanessa Berner4Sujata G. Pandit5Marcellene A. Gates-Hollingsworth6Kathryn J. Pflughoeft7Douglas C. Montgomery8Supriya Atta9Tuan Vo-Dinh10David P. AuCoin11Frederic Zenhausern12Center for Applied NanoBioscience and Medicine, College of Medicine, University of Arizona, Phoenix, Arizona, USADepartment of Microbiology and Immunology, University of Nevada Reno School of Medicine, Reno, Nevada, USACenter for Applied NanoBioscience and Medicine, College of Medicine, University of Arizona, Phoenix, Arizona, USACenter for Applied NanoBioscience and Medicine, College of Medicine, University of Arizona, Phoenix, Arizona, USADepartment of Microbiology and Immunology, University of Nevada Reno School of Medicine, Reno, Nevada, USADepartment of Microbiology and Immunology, University of Nevada Reno School of Medicine, Reno, Nevada, USADepartment of Microbiology and Immunology, University of Nevada Reno School of Medicine, Reno, Nevada, USADepartment of Microbiology and Immunology, University of Nevada Reno School of Medicine, Reno, Nevada, USASchool of Computing and Augmented Intelligence, Arizona State University, Tempe, Arizona, USAFitzpatrick Institute for Photonics, Departments of Biomedical Engineering, Duke University, Durham, North Carolina, USAFitzpatrick Institute for Photonics, Departments of Biomedical Engineering, Duke University, Durham, North Carolina, USADepartment of Microbiology and Immunology, University of Nevada Reno School of Medicine, Reno, Nevada, USACenter for Applied NanoBioscience and Medicine, College of Medicine, University of Arizona, Phoenix, Arizona, USAABSTRACT Rift Valley fever (RVF) is a vector-borne, zoonotic infectious disease with a proven history of morbidity and mortality in both humans and animals. Rift Valley fever virus (RVFV) is categorized as a high-priority biothreat agent by the Centers for Disease Control and Prevention and poses a serious national threat due to its ease of dissemination and potential for social disruption. RVF often presents as a febrile disease without specific symptoms, making early-stage detection particularly challenging. As such, it is critical that rapid, sensitive, and specific diagnostics are available for the detection of RVFV. While lateral flow immunoassays (LFIs) have been developed and validated for point-of-care (POC) diagnostics, vertical flow immunoassays (VFIs) provide enhanced analytical sensitivity and are equally suitable for POC use. In this study, we developed a VFI system for the detection of RVFV, achieving a limit of detection of 0.78 ng/mL, which is a 2.5-fold increase in analytical sensitivity compared to an LFI prototype. Furthermore, minimal cross-reactivity was demonstrated when performing the assay with target analytes of other high-priority biothreats and one other common viral nucleoprotein. This high-sensitivity VFI has the potential to prove useful for the detection of RVFV and other high-priority biothreat agents at the POC.IMPORTANCEIn this study, we have developed a rapid, sensitive vertical flow immunoassay (VFI) for the detection of Rift Valley fever virus (RVFV) in spiked human serum. The prototype diagnostic described in this research was shown to be more sensitive than traditional methods, such as lateral flow dipstick tests. Moreover, the VFI is readily deployable at the point of care in resource-limited settings. The ability of the described diagnostic to accurately and rapidly detect RVFV in samples could expedite the delivery of life-saving care and thus improve patient outcomes.https://journals.asm.org/doi/10.1128/spectrum.00341-25Rift Valley fever virusvertical flow devicebiothreat agentscolorimetric detectionrapid antigen test |
| spellingShingle | Alexander J. Summers Haydon J. Hill Jasmine P. Devadhasan Jian Gu Vanessa Berner Sujata G. Pandit Marcellene A. Gates-Hollingsworth Kathryn J. Pflughoeft Douglas C. Montgomery Supriya Atta Tuan Vo-Dinh David P. AuCoin Frederic Zenhausern Development of a high-sensitivity vertical flow immunoassay for the detection of Rift Valley fever virus Microbiology Spectrum Rift Valley fever virus vertical flow device biothreat agents colorimetric detection rapid antigen test |
| title | Development of a high-sensitivity vertical flow immunoassay for the detection of Rift Valley fever virus |
| title_full | Development of a high-sensitivity vertical flow immunoassay for the detection of Rift Valley fever virus |
| title_fullStr | Development of a high-sensitivity vertical flow immunoassay for the detection of Rift Valley fever virus |
| title_full_unstemmed | Development of a high-sensitivity vertical flow immunoassay for the detection of Rift Valley fever virus |
| title_short | Development of a high-sensitivity vertical flow immunoassay for the detection of Rift Valley fever virus |
| title_sort | development of a high sensitivity vertical flow immunoassay for the detection of rift valley fever virus |
| topic | Rift Valley fever virus vertical flow device biothreat agents colorimetric detection rapid antigen test |
| url | https://journals.asm.org/doi/10.1128/spectrum.00341-25 |
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