Analytical and clinical evaluation of a duplex RT-qPCR assay for the detection and identification of o'nyong-nyong and chikungunya virus.

Autor:	Wesselmann KM; Unité des Virus Émergents (UVE: Aix-Marseille Univ, Università di Corsica, IRD 190, Inserm 1207, IRBA), Marseille, France., Luciani L; Unité des Virus Émergents (UVE: Aix-Marseille Univ, Università di Corsica, IRD 190, Inserm 1207, IRBA), Marseille, France.; Assistance publique-hôpitaux de Marseille (AP-HM), Service de virologie aiguë et tropicale, Marseille, France., Thirion L; Unité des Virus Émergents (UVE: Aix-Marseille Univ, Università di Corsica, IRD 190, Inserm 1207, IRBA), Marseille, France., de Lamballerie X; Unité des Virus Émergents (UVE: Aix-Marseille Univ, Università di Corsica, IRD 190, Inserm 1207, IRBA), Marseille, France.; Assistance publique-hôpitaux de Marseille (AP-HM), Service de virologie aiguë et tropicale, Marseille, France.; Centre National de Référence des Arbovirus, Inserm-IRBA, Marseille, France., Charrel R; Unité des Virus Émergents (UVE: Aix-Marseille Univ, Università di Corsica, IRD 190, Inserm 1207, IRBA), Marseille, France.; Assistance publique-hôpitaux de Marseille (AP-HM), Service de virologie aiguë et tropicale, Marseille, France., Pezzi L; Unité des Virus Émergents (UVE: Aix-Marseille Univ, Università di Corsica, IRD 190, Inserm 1207, IRBA), Marseille, France.; Centre National de Référence des Arbovirus, Inserm-IRBA, Marseille, France.
Jazyk:	angličtina
Zdroj:	Emerging microbes & infections [Emerg Microbes Infect] 2024 Dec; Vol. 13 (1), pp. 2429650. Date of Electronic Publication: 2024 Nov 24.
DOI:	10.1080/22221751.2024.2429650
Abstrakt:	The mosquito-borne alphavirus o'nyong-nyong virus (ONNV) has proven its potential to cause major human outbreaks. On the African continent, ONNV causes unspecific febrile illness and co-circulates with the close relative chikungunya virus (CHIKV). The true scale of ONNV burden is poorly understood in Africa, because of the scarce availability of molecular in-house and commercial assays, strong cross-reactivity between ONNV and CHIKV in serological assays and a lack of surveillance. We designed a new RT-qPCR assay targeting the E1 gene for the detection of ONNV that can be used in monoplex or in duplex format, combined with a previously published CHIKV monoplex assay targeting the nsp1. The lower limit of detection with 95% positivity rate was determined to be <10 RNA copies/µL in monoplex and duplex format for both ONNV and CHIKV. Both monoplex assays and the duplex assay proved to be linear within the tested range of approximately 10 8 to 10 2 RNA copies/µl, and showed 100% specificity against a wide panel of arbovirus supernatants as well as several other pathogens in clinical samples. Testing of CHIKV positive serum and ONNV-spiked plasma samples confirmed the suitability of the assays in a clinical setting. The new assays provide a robust tool for molecular ONNV as well as ONNV/CHIKV simultaneous detection and may contribute to clarify the true burden of the two viruses, to improve arbovirus surveillance and to strengthen epidemics preparedness in Africa.
Databáze:	MEDLINE
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