Evaluation of a Trio Toscana Virus Real-Time RT-PCR Assay Targeting Three Genomic Regions within Nucleoprotein Gene

• Main Authors: Laurence THIRION, Laura PEZZI, Irene PEDROSA-CORRAL, Sara SANBONMATSU-GAMEZ, Xavier de LAMBALLERIE, Alessandra FALCHI, Mercedes PEREZ-RUIZ, Remi N. CHARREL
• Format: Article
• Language: English
• Published: MDPI AG 2021-02-01
• Series: Pathogens
• Subjects: <i>Phlebovirus</i>; <i>Phenuiviridae</i>; meningitis; sand fly; <i>Phlebotomus</i>; arbovirus
• Online Access: https://www.mdpi.com/2076-0817/10/3/254

Toscana virus (TOSV) can cause central nervous system infections in both residents of and travelers to Mediterranean countries. Data mining identified three real-time RT-qPCR assays for detecting TOSV RNA targeting non-overlapping regions in the nucleoprotein gene. Here, they were combined to create a multi-region assay named Trio TOSV RT-qPCR consisting of six primers and three probes. In this study, (i) we evaluated in silico the three RT-qPCR assays available in the literature for TOSV detection, (ii) we combined the three systems to create the Trio TOSV RT-qPCR, (iii) we assessed the specificity and sensitivity of the three monoplex assays versus the Trio TOSV RT-qPCR assay, and (iv) we compared the performance of the Trio TOSV RT-qPCR assay with one of the reference monoplex assays on clinical samples. In conclusion, the Trio TOSV RT-qPCR assay performs equally or better than the three monoplex assays; therefore, it provides a robust assay that can be used for both research and diagnostic purposes.