猪流行性腹泻病毒HB201602株的分离鉴定及S1基因序列分析

doi:10.14088/j.cnki.issn0439-8114.2019.24.055

湖北农业科学 ›› 2019, Vol. 58 ›› Issue (24): 227-231.doi: 10.14088/j.cnki.issn0439-8114.2019.24.055

猪流行性腹泻病毒HB201602株的分离鉴定及S1基因序列分析

刘威, 袁芳艳, 周丹娜, 刘泽文, 杨克礼, 段正赢, 郭锐, 高婷, 梁婉, 田永祥

湖北省农业科学院畜牧兽医研究所/农业农村部兽用药物与诊断技术湖北科学观测实验站,武汉 430064

收稿日期:2019-09-19 出版日期:2019-12-25 发布日期:2019-12-25
通讯作者: 田永祥,研究员,主要从事家禽疫病防控及兽用生物制品创制研究,（电话）027-87380195。
作者简介:刘威（1985-）,男,湖北武汉人,助理研究员,博士,主要从事动物传染病研究工作,（电话）027-87156079（电子信箱）liuwei85@126.com
基金资助:
国家重点研发计划项目（2016YFD0500906）; 湖北省农业科技创新中心资助项目（2015-620-004-001）

Isolation and identification of HB201602 strain of porcine epidemic diarrhea virus and phylogenetic tree analyses based on S1 gene

LIU Wei, YUAN Fang-yan, ZHOU Dan-na, LIU Ze-wen, YANG Ke-li, DUAN Zheng-ying, GUO Rui, GAO Ting, LIANG Wan, TIAN Yong-xiang

Institute of Animal Science and Veterinary Medicine,Hubei Academy of Agricultural Science/Hubei Science-observation Experiment Station of Veterinary Medicine and Diagnostic Technology （Ministry of Agriculture and Rural Affairs）,Wuhan 430064,China

Received:2019-09-19 Online:2019-12-25 Published:2019-12-25

摘要/Abstract

摘要： 从湖北地区规模化养殖场疑似腹泻病料中成功检测并分离获得了一株猪流行性腹泻病毒,分离毒株感染Vero细胞可引起明显的细胞病变,病毒效价达1.0×10^5.6 TCID₅₀/0.1 mL。间接免疫荧光结果显示HB201602感染细胞后,能被抗PEDV S蛋白的特异性单克隆抗体所识别,进一步证实所分离毒株为PEDV病毒。基于S1基因构建了系统发育树,表明2011年后分离的毒株大多以G2型变异毒株为主,HB201602属于G2-a亚型（变异毒株）簇。S1基因遗传变异分析发现G1-b亚型毒株与其他簇毒株相比存在9个特异的氨基酸突变,G1-a亚型经典毒株存在4个特异的氨基酸突变,S1基因的变异分析将有助于了解PEDV的遗传变异趋势,为新型疫苗的研发提供依据。

关键词: 猪流行性腹泻, 分离鉴定, 序列分析, 系统发育树, 遗传变异

Abstract: Porcine epidemic diarrhea virus （PEDV） HB201602 strain was isolated from the piglets suspected to be suffering a severe diarrhea in Hubei Province. The isolated strain can infect Vero cells and cause obvious cell pathological change, with viral titer up to 1.0×10^5.6 TCID₅₀/0.1ml. Indirect immunofluorescence results indicated that HB201602 infected cells could be recognized by monoclonal antibodies against PEDV S protein, further confirming that the isolated strain was PEDV virus. Phylogenetic trees were constructed based on S1 gene, and the results indicated that most of the strains isolated after 2011 were G2 variant strains, and HB201602 belonged to G2-a subtype （variant strain） cluster. Genetic variation analysis of S1 gene shows that G1-b subtype strain has 9 specific single amino acid mutations compared with other cluster strains, and G1-a subtype classical strain has 4 specific amino acid mutations. Variation analysis of S1 gene will provide a basis for understanding the genetic variation trend of PEDV and the development of new vaccines.

Key words: porcine epidemic diarrhea virus, isolation, sequence analysis, phylogenetic tree, genetic variation

中图分类号:

S852.65⁺1

刘威, 袁芳艳, 周丹娜, 刘泽文, 杨克礼, 段正赢, 郭锐, 高婷, 梁婉, 田永祥. 猪流行性腹泻病毒HB201602株的分离鉴定及S1基因序列分析[J]. 湖北农业科学, 2019, 58(24): 227-231.

LIU Wei, YUAN Fang-yan, ZHOU Dan-na, LIU Ze-wen, YANG Ke-li, DUAN Zheng-ying, GUO Rui, GAO Ting, LIANG Wan, TIAN Yong-xiang. Isolation and identification of HB201602 strain of porcine epidemic diarrhea virus and phylogenetic tree analyses based on S1 gene[J]. HUBEI AGRICULTURAL SCIENCES, 2019, 58(24): 227-231.

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猪流行性腹泻病毒HB201602株的分离鉴定及S1基因序列分析

Isolation and identification of HB201602 strain of porcine epidemic diarrhea virus and phylogenetic tree analyses based on S1 gene

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