一株牛口咽液分离口蹄疫病毒的特性分析
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作者单位:

1.中国农业科学院兰州兽医研究所/兰州大学 动物医学与生物安全学院,动物疫病防控全国重点实验室,甘肃 兰州;2.WOAH/国家口蹄疫参考实验室,甘肃 兰州;3.华中农业大学 动物医学院,湖北 武汉;4.甘肃省病原生物学基础学科研究中心,甘肃 兰州

作者简介:

蔡梦婷:收集样品,操作试验;袁红:操作试验,修改文章;王利豪:操作试验;杨竞纯:操作试验;王涛:分析数据;包慧芳:指导试验操作;杨宇轩:操作试验;孙普:分析数据;李坤:指导试验操作;黄书伦:操作试验;郭海洋:辅助试验操作;李平花:分析数据;卢曾军:修改文章;何继军:设计试验,撰写文章;白兴文:设计试验,撰写文章。

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基金项目:

国家重点研发计划(2021YFD1800300);甘肃省科技计划(23JRRA1515);甘肃省重大科技专项(21ZD3NA001, 22ZD6NA001, 22ZD6NA012);国家生猪技术创新中心项目(NCTIP-XD/C03)


Characteristics of a strain of foot-and-mouth disease virus isolated from bovine oropharyngeal fluid
Author:
Affiliation:

1.State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China;2.World Organization for Animal Health (WOAH)/National Foot and Mouth Disease Reference Laboratory, Lanzhou, Gansu, China;3.College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, China;4.Gansu Province Research Center for Basic Disciplines of Pathogen Biology, Lanzhou, Gansu, China

Fund Project:

This work was supported by the National Key Research and Development Program of China (2021YFD1800300), the Science and Technology Plan of Gansu Province (23JRRA1515), the Major Science and Technology Project of Gansu Province (21ZD3NA001, 22ZD6NA001, 22ZD6NA012), and the Project of National Center of Technology Innovation for Pigs (NCTIP-XD/C03).

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    摘要:

    目的 分离、鉴定一株牛源口咽液中的口蹄疫病毒(foot-and-mouth disease virus, FMDV),分析其抗原蛋白氨基酸变异及增殖特性。方法 采用RT-PCR及基因测序技术鉴定FMDV阳性牛口咽液样本,通过细胞接种从阳性样本中分离病毒,并利用间接免疫荧光法对分离病毒进行鉴定。通过PCR分段扩增及序列测定获得分离FMDV的全基因组,进一步通过序列比对分析分离FMDV的抗原蛋白氨基酸变异,通过蚀斑形成试验和一步生长曲线评价分离FMDV的增殖特性。结果 成功从牛口咽液中分离、鉴定出一株FMDV,命名为O/FMDV/OP/2022/B。该病毒属于O型Ind-2001谱系,其抗原蛋白VP1、VP2和VP3的氨基酸残基发生变异。在BHK-21细胞上,O/FMDV/OP/2022/B毒株形成的蚀斑显著小于疫苗毒株O/FMDV/HN/93的蚀斑,且其一步生长曲线明显低于O/FMDV/HN/93。结论 本研究从牛口咽液中分离出一株FMDV,发现其抗原蛋白氨基酸发生变异,且增殖能力显著低于疫苗毒株O/FMDV/HN/93。该研究有助于从病毒学角度深入理解FMDV持续感染牛口咽组织的机制,并为口蹄疫的防控提供了参考依据。

    Abstract:

    Objective To isolate and identify a strain of foot-and-mouth disease virus (FMDV) from bovine oropharyngeal fluid (OPF) and subsequently analyze the amino acid mutations in antigenic proteins and the proliferative properties of the isolated virus.Methods RT-PCR and sequencing were performed to identify the FMDV-positive OPF samples. The FMDV strain was isolated from bovine OPF samples by cell inoculation and identified by indirect immunofluorescence assay. The whole genome sequence of the strain was obtained by fragment amplification and sequencing. The amino acid sequences were compared to reveal the amino acid mutations in the antigenic proteins of the strain. The proliferative properties of the isolated FMDV strain were analyzed by the plaque forming assay and one-step growth curve.Results A FMDV strain O/FMDV/OP/2022/B was successfully isolated from bovine OPF, belonging to serotype O and Ind-2001 lineage. Amino acid mutations were identified in the antigenic proteins VP1, VP2, and VP3 of O/FMDV/OP/2022/B. In addition, the plaque of O/FMDV/OP/2022/B in BHK-21 cells was smaller than that of the vaccine strain O/FMDV/HN/93. The one-step growth curves showed that O/FMDV/OP/2022/B had significantly lower replication titer than O/FMDV/HN/93 in BHK-21 cells.Conclusion A strain of FMDV was isolated from bovine OPF, and it displayed amino acid mutations in antigenic proteins and had lower proliferative capacity than the vaccine strain O/FMDV/HN/93. This study contributes to a better understanding of the mechanism of persistent FMDV infection in the bovine oropharynx and provides a reference for the prevention and control of FMD.

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蔡梦婷,袁红,王利豪,杨竞纯,王涛,包慧芳,杨宇轩,孙普,李坤,黄书伦,郭海洋,李平花,卢曾军,何继军,白兴文. 一株牛口咽液分离口蹄疫病毒的特性分析[J]. 微生物学报, 2025, 65(7): 2938-2947

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  • 收稿日期:2024-12-12
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  • 在线发布日期: 2025-07-04
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