猪流行性腹泻病毒HBsh-1的分离鉴定和遗传演化分析  

Isolation,Identification and Genetic Evolution Analysis of Porcine Epidemic Diarrhea Virus HBsh-1

作　　者：王迪 陈冬杰 赵鸿远 WANG Di;CHEN Dongjie;ZHAO Hongyuan(School of Agroforestry and Medicine,Open University of China,Beijing 100039,China;The Institute of Animal Quarantine,Chinese Academy of Inspection and Quarantine,Beijing 100176,China;School of Modern Agriculture&Biotechnology,Ankang University,Ankang 725000,China)

机构地区：[1]国家开放大学农林医药教学部,北京100093 [2]中国检验检疫科学研究院动物检验与检疫研究所,北京100176 [3]安康学院现代农业与生物科技学院,安康725000

出　　处：《中国动物传染病学报》2023年第2期93-100,共8页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然科学基金青年项目(31902302);陕西省大学生创新创业训练计划项目(S202011397031);安康学院科技计划项目(2019AYQJ11)。

摘　　要：为了解河北地区猪流行性腹泻流行现状,将河北某规模化养猪场获得的腹泻病料处理后接种Vero CCL81细胞,盲传7代,各代次感染细胞裂解液中均可扩增阳性条带。经过噬斑纯化后,获得1株Vero CCL81细胞适应性良好的猪流行性腹泻病毒(PEDV)毒株,命名为HBsh-1。间接免疫荧光结果显示HBsh-1感染细胞后,能被PEDV N蛋白单克隆抗体特异性识别。分片段扩增HBsh-1的S基因后测定其序列。S基因序列分析结果表明,HBsh-1与经典毒株CV777的S基因核苷酸相似性为94.1%,氨基酸相似性为93.3%。与经典毒株相比,S基因存在4处连续核苷酸插入或缺失。S基因遗传演化分析结果显示,HBsh-1与PEDV变异毒株亲缘关系较近,属于GII-b亚型。以上研究结果表明,我们成功分离得到1株归属于GII-b亚型的PEDV新毒株,为研究分析当地PEDV流行现状和演化趋势提供依据,为制定猪流行性腹泻疫情防控策略提供参考。To study the epidemic situation of porcine epidemic diarrhea(PED)in Hebei province,the diarrheal materials obtained from a large-scale pig farm in Hebei province were inoculated onto Vero CCL81 cells for blind passages,and the positive bands were amplified from the lysates of infected cells for each passage.After plaque purification,a PEDV with good adaptability of Vero CCL81 cells was obtained and designated as HBsh-1 strain.Indirect immunofluorescence results showed that HBsh-1 infected cells were specifically recognized by the monoclonal antibody specific for PEDV N protein.The S gene of HBsh-1 was amplified by fragment and sequenced.The homology of S gene nucleotide and amino acid between HBsh-1 and CV777 strain was 94.1%and 93.3%respectively.As compared with the classical strain,S gene had 4 successive nucleotide insertions and deletions.Genetic evolution analysis of S gene showed that HBsh-1 belonged to the GII-b subtype and was closely related to PEDV mutant strain.The above results indicated that a new PEDV strain belonging to the GII-b subtype was successfully isolated,which provided a basis for the analysis of the epidemic status and evolutionary trend of local PEDV and a reference for the development of PED prevention and control strategies.

关 键 词：猪流行性腹泻病毒 S基因 分离 鉴定 遗传演化 

分 类 号：S852.651[农业科学—基础兽医学]