Laboratório de Virologia, Programa de Pós-Graduação em Ciências da Saúde, Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso, 78060-900, Brazil.
Laboratório de Proteção Florestal, Programa de Pós-Graduação em Ciências Florestais e Ambientais, Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso, 78060-900, Brazil.
Parasit Vectors. 2018 Jul 11;11(1):405. doi: 10.1186/s13071-018-2985-3.
High throughput sequencing (HTS) boosted the discovery of novel viruses and new variants of known viruses. Here we investigated the presence of viruses in 12 pools of sand flies captured in three climatic periods in RAPELD grids at Rio Claro, Chapada dos Guimarães and at Pirizal, North Pantanal, Mato Grosso State, Midwestern Brazil by HTS, viral isolation of a putative Phlebovirus positive pool in Vero cells, RT-PCR and transmission electron microscopy (TEM).
One pool containing three Lutzomyia (Lutzomyia) longipalpis sand flies captured in the transitional climatic period in North Pantanal showed a tripartite genomic sequence of a putative novel Phlebovirus belonging to the phlebotomus fever serogroup. Phylogenetic analysis revealed this virus is closely related and share a common ancestor with phleboviruses included in the same clade: Chagres, Urucuri and Uriurana virus. RNA-dependent RNA polymerase (RdRP) presented 60%, 59% and 58% of amino-acid (aa) similarity with these phleboviruses, respectively. Similarity of Nucleoprotein and NSs protein codified by ambissense strategy of segment S was of 49% and 37%, respectively, with the proteins of the closest phlebovirus, Uriurana virus. Glycoproteins (G1, G2) and NSm protein presented 49% and 48% aa similarity with Chagres and Uriurana virus, respectively. Uriurana virus was isolated from sand flies in Brazilian Amazon and Urucuri from rodents in Utinga forest, Pará State. Chagres virus is an arbovirus responsible for outbreaks of febrile illness in Panama. This phlebovirus was isolated in Vero cells, confirmed by TEM and RT-PCR for the L segment of the virus, and named Viola phlebovirus.
HTS, viral isolation, RT-PCR and TEM showed the presence of one virus in sand flies from North Pantanal with identity to a putative novel Phlebovirus from phlebotomus fever serogroup, named Viola phlebovirus.
高通量测序（HTS）促进了新型病毒和已知病毒新变体的发现。在这里，我们通过 HTS 调查了巴西中西部马托格罗索州里奥克拉鲁、瓜马拉斯高地的 RAPELD 网格和北潘塔纳尔的皮里泽尔三个气候期采集的 12 个沙蝇样本中病毒的存在情况，对一个疑似黄病毒阳性样本进行了病毒分离，用 Vero 细胞进行 RT-PCR 和透射电子显微镜（TEM）检测。
北潘塔纳尔过渡气候期采集的三只有足按蚊沙蝇样本中发现了一个可能的新型黄病毒的三分体基因组序列，该病毒属于黄病毒属发热血清群。系统发育分析表明，该病毒与包括在同一分支内的黄病毒密切相关，并与 Chagres、Urucuri 和 Uriurana 病毒有共同的祖先。RNA 依赖性 RNA 聚合酶（RdRP）与这些黄病毒的氨基酸（aa）相似性分别为 60%、59%和 58%。由节段 S 的双义策略编码的核蛋白和 NSs 蛋白的相似性分别为 49%和 37%，与最接近的黄病毒 Uriurana 病毒的蛋白相似性。糖蛋白（G1、G2）和 NSm 蛋白与 Chagres 和 Uriurana 病毒的 aa 相似性分别为 49%和 48%。Uriurana 病毒是从巴西亚马逊地区的沙蝇中分离出来的，Urucuri 病毒是从帕拉州乌廷加森林的啮齿动物中分离出来的。Chagres 病毒是一种引起巴拿马发热疾病爆发的虫媒病毒。该黄病毒在 Vero 细胞中分离出来，通过 TEM 和 RT-PCR 对病毒的 L 节段进行了确认，并命名为 Viola 黄病毒。
HTS、病毒分离、RT-PCR 和 TEM 显示，北潘塔纳尔的沙蝇中存在一种与黄病毒发热血清群中的一种新型黄病毒具有相同身份的病毒，被命名为 Viola 黄病毒。
