在莫桑比克库蚊属(Culex spp.)蚊虫中发现一种新型微小 RNA 病毒的遗传特征。
Genetic characterization of a novel picorna-like virus in Culex spp. mosquitoes from Mozambique.
机构信息
Section of Virology, Department of Biomedical and Veterinary Public Health, Box 7028, Swedish University of Agricultural Sciences (SLU), 75007, Uppsala, Sweden.
SLU Global Bioinformatics Centre, Department of Animal Breeding and Genetics, Box 7023, Swedish University of Agricultural Sciences, Uppsala, Sweden.
出版信息
Virol J. 2018 Apr 18;15(1):71. doi: 10.1186/s12985-018-0981-z.
BACKGROUND
Mosquitoes are the potential vectors for a variety of viruses that can cause diseases in the human and animal populations. Viruses in the order Picornavirales infect a broad range of hosts, including mosquitoes. In this study, we aimed to characterize a novel picorna-like virus from the Culex spp. of mosquitoes from the Zambezi Valley of Mozambique.
METHODS
The extracted RNA from mosquito pools was pre-amplified with the sequence independent single primer amplification (SISPA) method and subjected to high-throughput sequencing using the Ion Torrent platform. Reads that are classified as Iflaviridae, Picornaviridae and Dicistroviridae were assembled by CodonCode Aligner and SPAdes. Gaps between the viral contigs were sequenced by PCR. The genomic ends were analyzed by 5' and 3' RACE PCRs. The ORF was predicted with the NCBI ORF finder. The conserved domains were identified with ClustalW multiple sequence alignment, and a phylogenetic tree was built with MEGA. The presence of the virus in individual mosquito pools was detected by RT-PCR assay.
RESULTS
A near full-length viral genome (9740 nt) was obtained in Culex mosquitoes that encoded a complete ORF (3112 aa), named Culex picorna-like virus (CuPV-1). The predicted ORF had 38% similarity to the Hubei picorna-like virus 35. The sequence of the conserved domains, Helicase-Protease-RNA-dependent RNA polymerase, were identified by multiple sequence alignment and found to be at the 3' end, similar to iflaviruses. Phylogenetic analysis of the putative RdRP amino acid sequences indicated that the virus clustered with members of the Iflaviridae family. CuPV-1 was detected in both Culex and Mansonia individual pools with low infection rates.
CONCLUSIONS
The study reported a highly divergent, near full-length picorna-like virus genome from Culex spp. mosquitoes from Mozambique. The discovery and characterization of novel viruses in mosquitoes is an initial step, which will provide insights into mosquito-virus interaction mechanisms, genetic diversity and evolution.
背景
蚊子是多种病毒的潜在载体，这些病毒会导致人类和动物群体患病。小 RNA 病毒目（Picornavirales）的病毒感染范围广泛，包括蚊子。在这项研究中，我们旨在从莫桑比克赞比西河谷的库蚊（Culex spp.）中鉴定一种新型的类微小 RNA 病毒。
方法
采用序列非依赖性单引物扩增（SISPA）方法对蚊子池的提取 RNA 进行预扩增，然后使用 Ion Torrent 平台进行高通量测序。使用 CodonCode Aligner 和 SPAdes 对分类为 Iflaviridae、Picornaviridae 和 Dicistroviridae 的读取进行组装。通过 PCR 测序病毒重叠序列之间的缺口。通过 5'和 3'RACE PCR 分析基因组末端。使用 NCBI ORF finder 预测 ORF。使用 ClustalW 多重序列比对鉴定保守结构域，并使用 MEGA 构建系统发育树。通过 RT-PCR 检测单个蚊子池中病毒的存在。
结果
从库蚊中获得了近全长病毒基因组（9740nt），该基因组编码一个完整的 ORF（3112aa），命名为库蚊微小 RNA 样病毒（CuPV-1）。预测的 ORF 与湖北微小 RNA 样病毒 35 具有 38%的相似性。通过多重序列比对鉴定了保守结构域（Helicase-Protease-RNA-dependent RNA polymerase），发现其位于 3'端，类似于 iflaviruses。推定 RdRP 氨基酸序列的系统发育分析表明，该病毒与 Iflaviridae 家族的成员聚类。CuPV-1 在库蚊和曼蚊个体池中有低感染率。
结论
本研究从莫桑比克库蚊中报道了一种高度分化的近全长微小 RNA 样病毒基因组。在蚊子中发现和鉴定新型病毒是一个初步步骤，这将为蚊子-病毒相互作用机制、遗传多样性和进化提供深入了解。
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