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Article

Real-Time Polymerase Chain Reaction Systems for Detection and Differentiation of Unclassified Viruses of the Phenuiviridae Family

by
Alena V. Dereventsova
1,*,
Alexander S. Klimentov
1,
Ivan S. Kholodilov
2,
Oxana A. Belova
2,
Alexander M. Butenko
3 and
Galina G. Karganova
2,4
1
Laboratory of Biochemistry, Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of RAS (Institute of Poliomyelitis), 108819 Moscow, Russia
2
Laboratory of Biology of Arboviruses, Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of RAS (Institute of Poliomyelitis), 108819 Moscow, Russia
3
D.I. Ivanovsky Institute of Virology Division of N.F. Gamaleya National Research Center of Epidemiology and Microbiology of the Ministry of Health of the Russian Federation, 1230986 Moscow, Russia
4
Institute for Translational Medicine and Biotechnology, Sechenov University, 119991 Moscow, Russia
*
Author to whom correspondence should be addressed.
Methods Protoc. 2025, 8(1), 20; https://doi.org/10.3390/mps8010020
Submission received: 13 December 2024 / Revised: 8 February 2025 / Accepted: 11 February 2025 / Published: 18 February 2025
(This article belongs to the Section Molecular and Cellular Biology)
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Abstract

The family Phenuiviridae, part of the order Hareavirales, includes arboviruses and arthropod-associated viruses, with sandflies, mosquitoes, and ticks as primary vectors. Historically, only sandfly/mosquito-borne phenuiviruses were associated with human diseases, but the emergence of severe fever with thrombocytopenia syndrome (SFTS) has highlighted the potential of tick-borne phenuiviruses as human pathogens. Recent discoveries of new arthropod-associated viruses, some of which remain unclassified, underscore the need for sensitive detection and differentiation methods, particularly in regions where these viruses may co-circulate. This study aimed to develop real-time PCR test systems for identifying and differentiating unclassified viruses within the Phenuiviridae family. In this study, tick suspensions containing phenuiviruses, previously obtained during the screening of ticks from various regions of Russia using pan-phenuivirus primers, were used. Specific primers and probes were designed to differentiate five Phenuiviridae viruses of genera Uukuvirus, Ixovirus, Phlebovirus and one unclassified phenuivirus, and their analytical sensitivity and specificity were evaluated. These PCR-based tools provide a robust method for detecting and classifying uncharacterized phenuiviruses, contributing to improved surveillance and understanding their potential epidemiological and epizootological impacts.
Keywords: Phenuiviridae; RT-PCR; tick-borne viruses; Uukuvirus; Ixovirus; Phlebovirus; Stavropol virus; Pedaselga virus; Kizhi virus; Gomselga virus Phenuiviridae; RT-PCR; tick-borne viruses; Uukuvirus; Ixovirus; Phlebovirus; Stavropol virus; Pedaselga virus; Kizhi virus; Gomselga virus

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MDPI and ACS Style

Dereventsova, A.V.; Klimentov, A.S.; Kholodilov, I.S.; Belova, O.A.; Butenko, A.M.; Karganova, G.G. Real-Time Polymerase Chain Reaction Systems for Detection and Differentiation of Unclassified Viruses of the Phenuiviridae Family. Methods Protoc. 2025, 8, 20. https://doi.org/10.3390/mps8010020

AMA Style

Dereventsova AV, Klimentov AS, Kholodilov IS, Belova OA, Butenko AM, Karganova GG. Real-Time Polymerase Chain Reaction Systems for Detection and Differentiation of Unclassified Viruses of the Phenuiviridae Family. Methods and Protocols. 2025; 8(1):20. https://doi.org/10.3390/mps8010020

Chicago/Turabian Style

Dereventsova, Alena V., Alexander S. Klimentov, Ivan S. Kholodilov, Oxana A. Belova, Alexander M. Butenko, and Galina G. Karganova. 2025. "Real-Time Polymerase Chain Reaction Systems for Detection and Differentiation of Unclassified Viruses of the Phenuiviridae Family" Methods and Protocols 8, no. 1: 20. https://doi.org/10.3390/mps8010020

APA Style

Dereventsova, A. V., Klimentov, A. S., Kholodilov, I. S., Belova, O. A., Butenko, A. M., & Karganova, G. G. (2025). Real-Time Polymerase Chain Reaction Systems for Detection and Differentiation of Unclassified Viruses of the Phenuiviridae Family. Methods and Protocols, 8(1), 20. https://doi.org/10.3390/mps8010020

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