Rapid identification of known and new RNA viruses from animal tissues.
Rapid identification of known and new RNA viruses from animal tissues.
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DOI：
10.1371/journal.ppat.1000163
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发表时间：
2008-09-26
期刊：
影响因子：
6.7
通讯作者：
Delwart EL
中科院分区：
文献类型：
--
作者：
Victoria JG;Kapoor A;Dupuis K;Schnurr DP;Delwart EL
病毒监测项目或诊断实验室偶尔会获得无法通过现有的细胞培养、血清学或核酸测试进行分类的感染样本。从1955年至1980年间收集的五个这样的样本，来自昆虫池、臭鼬脑、人类粪便和下水道污水，当接种到乳鼠身上时，导致了病理。在这项研究中，部分纯化的病毒核酸的序列独立扩增和小规模鸟枪测序在小鼠的脑和肌肉组织中得到了应用。在每个样本中都发现了一种单一的病毒病原体。对于每种病毒，仅通过对42-108个质粒插入进行测序就获得了16%至57%的病毒基因组。来源于人类排泄物或下水道污水的病毒属于微小核糖核病毒科，与已知的微小核糖核酸病毒具有80%至91%的氨基酸同源性。其中一种病毒的完整多蛋白序列与暂定的萨佩尔病毒属中的猴小核糖核酸病毒序列高度相似。昆虫和臭鼬来源的病毒序列与呼肠孤病毒科病毒片段基因组的氨基酸同源性在25%到98%之间。两个分离株高度不同：一个可能是正病毒属的一个新物种，另一个是环状病毒属的一个新物种。我们证明了一种简单、廉价和快速的元基因组学方法可以有效地在急性感染的小鼠的匀浆组织中识别已知的和高度分化的新病毒。病毒监测计划旨在识别正在传播的病毒，以保护公众和牲畜免受病毒爆发的影响。偶尔，怀疑携带病毒的样本会导致实验室动物患上严重疾病，但研究人员无法确定病毒的身份。在这里，我们应用了一种简单的病毒发现技术来直接从接种有症状的小鼠的组织中识别病毒，并使用快速病毒颗粒纯化和随机核酸扩增的方法在每个样本中发现了一种病毒。有两种病毒似乎与PicorNaviridae家族的成员关系密切。在另外三个样本中，最初从被压碎的蚊子池和一只患病臭鼬的大脑中收集，发现了一种与呼肠孤病毒科成员有关的已知和新病毒。呼肠孤病毒被认为是家畜的主要病原体。我们的研究为进一步分析这些不同病毒的流行和发病机制提供了基础，并说明了在急性感染动物的组织中可以很容易地识别出新的病毒种类。
Viral surveillance programs or diagnostic labs occasionally obtain infectious samples that fail to be typed by available cell culture, serological, or nucleic acid tests. Five such samples, originating from insect pools, skunk brain, human feces and sewer effluent, collected between 1955 and 1980, resulted in pathology when inoculated into suckling mice. In this study, sequence-independent amplification of partially purified viral nucleic acids and small scale shotgun sequencing was used on mouse brain and muscle tissues. A single viral agent was identified in each sample. For each virus, between 16% to 57% of the viral genome was acquired by sequencing only 42–108 plasmid inserts. Viruses derived from human feces or sewer effluent belonged to the Picornaviridae family and showed between 80% to 91% amino acid identities to known picornaviruses. The complete polyprotein sequence of one virus showed strong similarity to a simian picornavirus sequence in the provisional Sapelovirus genus. Insects and skunk derived viral sequences exhibited amino acid identities ranging from 25% to 98% to the segmented genomes of viruses within the Reoviridae family. Two isolates were highly divergent: one is potentially a new species within the orthoreovirus genus, and the other is a new species within the orbivirus genus. We demonstrate that a simple, inexpensive, and rapid metagenomics approach is effective for identifying known and highly divergent new viruses in homogenized tissues of acutely infected mice. Viral surveillance programs aim to identify circulating viruses to safeguard the public and livestock from viral outbreaks. Occasionally, samples suspected of harboring a virus cause severe disease in laboratory animals, but the identity of the virus eludes researchers. Here, we applied a simple viral discovery technique to identify viruses directly from the tissues of inoculated symptomatic mice and found a single virus in each sample using a rapid viral particle purification and random nucleic acid amplification method. Two viruses appear to be closely related to the members of the Picornaviridae family. In three other samples, originally collected from pools of crushed mosquitoes and the brain of a sick skunk, a known and novel viruses related to members of the Reoviridiae family were identified. Reoviruses are considered major pathogens of livestock. Our studies provide the groundwork for further analysis of the prevalence and pathogenesis of these divergent viruses and illustrate the ease with which new viral species can be identified in tissues of acutely infected animals.
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影响因子：
5.4
作者：
Limn, CK;Staeuber, N;Roy, P
通讯作者：
Roy, P
影响因子：
6.4
作者：
Excoffon, Katherine J. D. A.;Guglielmi, Kristen M.;Zabner, Joseph
通讯作者：
Zabner, Joseph
影响因子：
2.7
作者：
Granerod, Julia;Crowcroft, Natasha S.
通讯作者：
Crowcroft, Natasha S.
DOI：
10.1073/pnas.0504666102
发表时间：
2005-09-06
影响因子：
11.1
作者：
Allander, T;Tammi, MT;Andersson, B
通讯作者：
Andersson, B
影响因子：
3.8
作者：
Biagini, Philippe;Uch, Rathviro;de Micco, Philippe
通讯作者：
de Micco, Philippe