Thirion, LaurencePezzi, LauraPedrosa-Corral, IreneSanbonmatsu-Gamez, Sarade Lamballerie, XavierFalchi, AlessandraPerez-Ruiz, MercedesCharrel, Remi N2025-01-072025-01-072021-02-242076-0817https://hdl.handle.net/10668/28177Toscana 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.enAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/MediterraneanPhenuiviridaePhlebotomusPhlebovirusarbovirusdiagnosticsmeningitissand flyresearch article33668339open access10.3390/pathogens10030254PMC7996202https://www.mdpi.com/2076-0817/10/3/254/pdf?version=1615340154https://pmc.ncbi.nlm.nih.gov/articles/PMC7996202/pdf