Brisbarre Nadège, Plumet Sébastien, Cotteaux-Lautard Christelle, Emonet Sébastien F, Pages Frédéric, Leparc-Goffart Isabelle
UMR 7268 ADES, Aix-Marseille Université/EFS/CNRS, 27 Boulevard Jean Moulin, 13385 Marseille Cedex 05, France.
CNR Arbovirus IRBA (Institut de Recherche Biomédicale des Armées), 111 Avenue de la Corse BP40026, 13568 Marseille Cedex 02, France.
J Clin Virol. 2015 May;66:107-11. doi: 10.1016/j.jcv.2015.03.007. Epub 2015 Mar 10.
To scan a virus (TOSV) belongs to the Phlebovirus genus within the Bunyaviridae family. TOSV is an arbovirus transmitted by sandflies. In Mediterranean countries, TOSV is one of the major viral pathogens involved in aseptic meningitis and meningoencephalitis.
Development and assessment of a new sensitive and specific real-time RT-PCR assay for TOSV diagnosis.
TOSV-specific primers and probe targeting the S-segment of the genome were designed, based on recent TOSV sequences available in public databases. Sensitivity was assessed using 10-fold serial dilutions of a RNA transcript and serial dilutions of TOSV strains isolated from infected human beings. Specificity was determined by testing RNA extracts from closely related Phleboviruses. The assay was then used for TOSV infection diagnosis in 971 clinical samples and for TOSV detection in 2000 sandflies.
The real-time RT-PCR assay exhibited a sensitivity of under 257 copies per reaction for the RNA transcripts and 0.0056 and 0.014 TCID50 of Italian and Spanish TOSV genotypes per reaction, respectively. No other close Phleboviruses were detected. TOSV was identified in 17 clinical samples and in 3 sandflies.
The assay described is a rapid, robust and reliable real-time RT-PCR test for accurate diagnosis of human TOSV infection as well as for the surveillance of TOSV in vector populations.
托斯卡纳病毒（TOSV）属于布尼亚病毒科白蛉病毒属。TOSV是一种由白蛉传播的虫媒病毒。在地中海国家，TOSV是无菌性脑膜炎和脑膜脑炎的主要病毒病原体之一。
开发并评估一种用于TOSV诊断的新型灵敏且特异的实时逆转录聚合酶链反应（RT-PCR）检测方法。
基于公共数据库中最新的TOSV序列，设计了靶向基因组S片段的TOSV特异性引物和探针。使用RNA转录本的10倍系列稀释液以及从感染人类中分离出的TOSV毒株系列稀释液评估灵敏度。通过检测密切相关白蛉病毒的RNA提取物来确定特异性。然后将该检测方法用于971份临床样本的TOSV感染诊断以及2000只白蛉的TOSV检测。
实时RT-PCR检测方法对RNA转录本的灵敏度为每个反应低于257个拷贝，对意大利和西班牙TOSV基因型的灵敏度分别为每个反应0.0056和0.014半数组织培养感染剂量（TCID50）。未检测到其他密切相关的白蛉病毒。在17份临床样本和3只白蛉中鉴定出TOSV。
所描述的检测方法是一种快速、稳健且可靠的实时RT-PCR检测方法，可用于准确诊断人类TOSV感染以及监测媒介种群中的TOSV。
