Genetic characterization of a novel picorna-like virus in Culex spp. mosquitoes from Mozambique.
Genetic characterization of a novel picorna-like virus in Culex spp. mosquitoes from Mozambique.
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DOI：
10.1186/s12985-018-0981-z
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发表时间：
2018-04-18
期刊：
影响因子：
4.8
通讯作者：
Blomström AL
中科院分区：
文献类型：
--
作者：
Cholleti H;Hayer J;Fafetine J;Berg M;Blomström AL
蚊子是多种病毒的潜在传播媒介，这些病毒可在人类和动物群体中引发疾病。小核糖核酸病毒目（Picornavirales）中的病毒感染多种宿主，包括蚊子。在本研究中，我们旨在对来自莫桑比克赞比西河谷库蚊属（Culex spp.）蚊子的一种新型类小核糖核酸病毒进行特征分析。
从蚊子混合样本中提取的RNA采用不依赖序列的单引物扩增（SISPA）方法进行预扩增，并使用离子激流（Ion Torrent）平台进行高通量测序。被归类为伊夫拉病毒科（Iflaviridae）、小核糖核酸病毒科（Picornaviridae）和双顺反子病毒科（Dicistroviridae）的读段通过CodonCode Aligner和SPAdes软件进行组装。病毒重叠群之间的缺口通过PCR进行测序。基因组末端通过5′和3′快速扩增cDNA末端（RACE）PCR进行分析。开放阅读框（ORF）通过美国国立生物技术信息中心（NCBI）的ORF查找工具进行预测。保守结构域通过ClustalW多序列比对进行鉴定，并使用MEGA软件构建系统发育树。通过逆转录PCR（RT - PCR）检测单个蚊子混合样本中病毒的存在情况。
在库蚊中获得了一个近全长的病毒基因组（9740个核苷酸），它编码一个完整的开放阅读框（3112个氨基酸），被命名为库蚊类小核糖核酸病毒（Culex picorna - like virus，CuPV - 1）。预测的开放阅读框与湖北类小核糖核酸病毒35有38%的相似性。通过多序列比对鉴定出保守结构域——解旋酶 - 蛋白酶 - 依赖RNA的RNA聚合酶的序列，发现其位于3′端，与伊夫拉病毒相似。对假定的依赖RNA的RNA聚合酶（RdRP）氨基酸序列进行的系统发育分析表明，该病毒与伊夫拉病毒科成员聚为一类。在库蚊和曼蚊（Mansonia）的单个混合样本中均检测到CuPV - 1，但感染率较低。
该研究报道了来自莫桑比克库蚊属蚊子的一种高度分化的近全长类小核糖核酸病毒基因组。蚊子中新型病毒的发现和特征分析是第一步，这将为了解蚊子 - 病毒相互作用机制、遗传多样性和进化提供见解。
本文的网络版（10.1186/s12985 - 018 - 0981 - z）包含补充材料，授权用户可获取。
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. 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. 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. 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. The online version of this article (10.1186/s12985-018-0981-z) contains supplementary material, which is available to authorized users.
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