Establishment of a reverse genetics system for avian rotavirus A strain PO-13
Establishment of a reverse genetics system for avian rotavirus A strain PO-13
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DOI：
10.1099/jgv.0.001760
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发表时间：
2022-01-01
影响因子：
3.8
通讯作者：
Ito, Naoto
中科院分区：
文献类型：
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作者：
Kanda, Marika;Fukuda, Saori;Ito, Naoto
禽轮状病毒A（Avian Rotavirus A，RVA）是引起雏禽腹泻的主要肠道病原之一。重要的是，一些G18 P基因型的禽类RVA毒株[17]可自然传播至哺乳动物物种并引起临床疾病，表明其对动物健康的潜在风险。虽然禽RVA病毒株致病机理的分子信息有助于评估其风险，但缺乏针对这些病毒株的反向遗传学（RG）系统阻碍了其致病机制的阐明。在这项研究中，我们的目的是建立禽G18 P [17]原型菌株PO-13的RG系统，该菌株于1983年在日本从鸽子中分离出来，并在实验中证明对乳鼠具有致病性。用表达该毒株11个基因组RNA片段的质粒转染，导致感染性病毒的拯救，其基因组上具有人工引入的遗传标记，表明成功建立了PO-13毒株的RG系统。重组菌株rPO- 13的生物学特性与野生型菌株wtPO- 13基本一致.值得注意的是，rPO- 13和wtPO- 13均以相似的效率诱导乳鼠腹泻。这表明RG系统将有助于从分子水平上阐明PO-13株的致病机理。这是第一次报告建立了一个RG系统的禽RVA株。
Avian rotavirus A (RVA) is one of major enteric pathogens that cause diarrhoea in young avian individuals. Importantly, some of the avian RVA strains of G18P[17] genotype are naturally transmitted to and cause clinical diseases in mammalian species, indicating their potential risks to animal health. Although molecular information on the pathogenesis by avian RVA strains will be useful for estimating their risks, the absence of a reverse genetics (RG) system for these strains has hindered the elucida-tion of their pathogenic mechanisms. In this study, we aimed to establish an RG system for the avian G18P[17] prototype strain PO -13, which was isolated from a pigeon in Japan in 1983 and was experimentally shown to be pathogenic in suckling mice. Transfection with plasmids expressing 11 genomic RNA segments of the strain resulted in rescue of the infectious virus with an artificially introduced genetic marker on its genome, indicating that an RG system for the PO -13 strain was successfully established. The rescued recombinant strain rPO- 13 had biological properties almost identical to those of its wild -type strain (wtPO- 13). Notably, both rPO- 13 and wtPO- 13 induced diarrhoea in suckling mice with similar efficiencies. It was thus demon-strated that the RG system will be useful for elucidating the pathogenic mechanisms of the PO -13 strain at the molecular level. This is the first report of the establishment of an RG system for an avian RVA strain.