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利用纳米孔技术对流行期间的犬瘟热病毒进行全基因组测序。

Complete genomic sequencing of canine distemper virus with nanopore technology during an epizootic event.

机构信息

National Laboratory of Virology, Szentágothai Research Centre, University of Pécs, Pécs, 7624, Hungary.

Institute of Biology, Faculty of Sciences, University of Pécs, Pécs, 7624, Hungary.

出版信息

Sci Rep. 2022 Mar 8;12(1):4116. doi: 10.1038/s41598-022-08183-3.

Abstract

Canine distemper virus (CDV) endangers a wide range of wild animal populations, can cross species barriers and therefore representing a significant conservational and animal health risk around the globe. During spring to autumn 2021, according to our current estimates a minimum of 50 red foxes (Vulpes vulpes) died of CDV in Hungary, with CDV lesions. Oral, nasal and rectal swab samples were RT-PCR screened for Canine Distemper Virus from red fox carcasses. To investigate in more detail the origins of these CDV strains, 19 complete genomes were sequenced with a pan-genotype CDV-specific amplicon-based sequencing method developed by our laboratory and optimized for the Oxford Nanopore Technologies platform. Phylogenetic analysis of the complete genomic sequences and separately the hemagglutinin gene sequences revealed the role of the Europe lineage of CDV as a causative agent for the current epizootic. Here we highlight the growing importance of fast developing rapid sequencing technologies to aid rapid response activities during epidemics or epizootic events. We also emphasize the urgent need for improved surveillance of CDV, considering the epizootic capability of enzootic strains as reported in the current study. For such future efforts, we provide a novel NGS protocol to facilitate future genomic surveillance studies.

摘要

犬瘟热病毒(CDV)危害着广泛的野生动物种群,能够跨越物种屏障,因此在全球范围内构成了重大的保护和动物健康风险。在 2021 年春季至秋季期间,根据我们目前的估计,匈牙利至少有 50 只红狐(Vulpes vulpes)死于 CDV,且出现 CDV 病变。我们从红狐尸体中采集了口腔、鼻腔和直肠拭子样本,使用 RT-PCR 进行犬瘟热病毒筛查。为了更详细地研究这些 CDV 毒株的起源,我们使用我们实验室开发的基于 pan-genotype CDV 特异性扩增子的测序方法对 19 个完整基因组进行了测序,并对牛津纳米孔技术平台进行了优化。对完整基因组序列和单独的血凝素基因序列进行的系统发育分析表明,CDV 的欧洲谱系是当前疫病的病原体。在此,我们强调了快速发展的快速测序技术在大流行或疫病爆发期间快速响应活动的重要性。我们还强调了迫切需要改进 CDV 的监测,考虑到本研究报告的地方性流行株的流行能力。对于未来的此类工作,我们提供了一种新的 NGS 方案,以促进未来的基因组监测研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/af00/8904823/fd76cc2fd12f/41598_2022_8183_Fig1_HTML.jpg

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