Thirion Laurence, Pezzi Laura, Pedrosa-Corral Irene, Sanbonmatsu-Gamez Sara, de Lamballerie Xavier, Falchi Alessandra, Perez-Ruiz Mercedes, Charrel Remi N
Unité des Virus Émergents (UVE: Aix-Marseille Univ-IRD 190-Inserm 1207-IHU Méditerranée Infection), 13005 Marseille.
UR7310, Laboratoire de Virologie, Université de Corse-Inserm, 20250 Corte, France.
Pathogens. 2021 Feb 24;10(3):254. doi: 10.3390/pathogens10030254.
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.
托斯卡纳病毒（TOSV）可导致地中海国家的居民和旅行者发生中枢神经系统感染。数据挖掘确定了三种用于检测TOSV RNA的实时RT-qPCR检测方法，它们靶向核蛋白基因中的非重叠区域。在此，将它们组合起来创建了一种名为三重TOSV RT-qPCR的多区域检测方法，该方法由六种引物和三种探针组成。在本研究中，（i）我们通过计算机模拟评估了文献中可用的三种用于TOSV检测的RT-qPCR检测方法，（ii）我们将这三种系统组合起来创建了三重TOSV RT-qPCR，（iii）我们评估了三种单重检测方法与三重TOSV RT-qPCR检测方法的特异性和灵敏度，以及（iv）我们将三重TOSV RT-qPCR检测方法与一种参考单重检测方法在临床样本上的性能进行了比较。总之，三重TOSV RT-qPCR检测方法的性能与三种单重检测方法相当或更好；因此，它提供了一种可靠的检测方法，可用于研究和诊断目的。
