Ferreira Daniela Duque, Cook Shelley, Lopes Ângela, de Matos António Pedro, Esteves Aida, Abecasis Ana, de Almeida António Paulo Gouveia, Piedade João, Parreira Ricardo
Unidade de Microbiologia Médica, Grupo de Virologia, Instituto de Higiene e Medicina Tropical (IHMT), Universidade Nova de Lisboa (UNL), Rua da Junqueira 100, 1349-008, Lisbon, Portugal.
Virus Genes. 2013 Dec;47(3):532-45. doi: 10.1007/s11262-013-0960-9. Epub 2013 Jul 23.
We describe the isolation and characterization of an insect-specific flavivirus (ISF) from Ochlerotatus caspius (Pallas, 1771) mosquitoes collected in southern Portugal. The RNA genome of this virus, tentatively designated OCFVPT, for O. caspius flavivirus from Portugal, encodes a polyprotein showing all the features expected for a flavivirus. As frequently observed for ISF, the viral genomes seems to encode a putative Fairly Interesting Flavivirus ORF (FIFO)-like product, the synthesis of which would occur as a result of a -1 translation frameshift event. OCFVPT was isolated in the C6/36 Stegomyia albopicta (= Aedes albopictus) cell line where it replicates rapidly, but failed to replicate in Vero cells in common with other ISFs. Unlike some of the latter, however, the OCFVPT genome does not seem to be integrated in the mosquito cells we tested. Phylogenetic analyses based on partial ISF NS5 nucleotide sequences placed OCFVPT among recently published viral strains documented from mosquitoes collected in the Iberian Peninsula, while analyses of ORF/E/NS3/or NS5 amino acid sequences cluster OCFVPT with HANKV (Hanko virus), an ISF recently isolated from O. caspius mosquitoes collected in Finland. Taking into account the genetic relatedness with this virus, OCFVPT is not expected to be overtly cytopathic to C6/36 cells. The cytopathic effects associated with its presence in culture supernatants are postulated to be the result of the replication of a co-isolated putative new Negev-like virus.
我们描述了从葡萄牙南部采集的环跗库蚊（Ochlerotatus caspius，帕拉斯，1771年）中分离出一种昆虫特异性黄病毒（ISF）并对其进行表征的过程。这种病毒的RNA基因组，暂定名为OCFVPT，即来自葡萄牙的环跗库蚊黄病毒，编码一种多聚蛋白，具有黄病毒预期的所有特征。正如ISF中常见的那样，病毒基因组似乎编码一种假定的类似“相当有趣的黄病毒开放阅读框（FIFO）”的产物，其合成是由 -1 翻译移码事件导致的。OCFVPT在白纹伊蚊（Stegomyia albopicta = 埃及伊蚊）的C6/36细胞系中分离得到，在该细胞系中它能快速复制，但与其他ISF一样，未能在Vero细胞中复制。然而，与其中一些不同的是，OCFVPT基因组似乎并未整合到我们测试的蚊子细胞中。基于部分ISF NS5核苷酸序列的系统发育分析将OCFVPT置于伊比利亚半岛采集的蚊子中最近发表的病毒株之间，而对开放阅读框/E/NS3/或NS5氨基酸序列的分析则将OCFVPT与HANKV（汉科病毒）聚类，HANKV是最近从芬兰采集的环跗库蚊中分离出的一种ISF。考虑到与这种病毒的遗传相关性，预计OCFVPT对C6/36细胞不会有明显的细胞病变作用。推测与其存在于培养上清液中相关的细胞病变效应是共分离出的一种假定的新内盖夫样病毒复制的结果。
