猪繁殖与呼吸综合征病毒JX14T2毒株的分离与鉴定

发布时间：2018-09-14 16:20

【摘要】：江西省某猪场育肥猪群出现高热、呼吸困难和死亡等症状,为确定疫病病原,本实验室从发病猪场随机采取20份病死猪血样和组织,用RT-PCR的方法进行病原检测。结果显示,19份临床样品呈现猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)阳性。随后对病死猪肺组织进行病毒分离,结果显示分离的第1代病毒感染后48 h即可出现典型的致细胞病变效应(cytopathic effect,CPE),对传至第5代的病毒进行RT-PCR检测和间接免疫荧光鉴定,结果证实我们分离获得的病毒为PRRSV,命名为JX14T2。将JX14T2 P5代次的病毒与高致病性PRRSV强、弱毒代表株进行生长特性比较分析,结果显示,JX14T2在Marc-145细胞上形成的细胞病变、病毒噬斑形态以及病毒增殖动态均与HP-PRRSV毒株相似。对JX14T2进行全长基因测序和序列比较分析,结果表明JX14T2基因组全长14 960 bp,属于北美型毒株,其NSP2除存在的1+29个氨基酸的缺失而外,在598~717 aa还存在120个氨基酸的缺失,与HP-PRRSV亲缘关系较近。本研究结果为掌握PRRSV流行情况以及致病机制提供了科学依据。
[Abstract]:High fever, dyspnea and death were found in fattening pigs at a pig farm in Jiangxi Province. In order to determine the pathogen of epidemic disease, 20 blood samples and tissues of dead pigs were randomly collected from the pig farm. The pathogen was detected by RT-PCR method. The results showed that 19 clinical samples were positive for porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus,PRRSV). The virus was isolated from the lung tissue of the dead pigs. The results showed that typical cytopathic effect (cytopathic effect,CPE) appeared 48 hours after the first generation virus was infected. The virus transmitted to the fifth generation was detected by RT-PCR and identified by indirect immunofluorescence. The results confirmed that the virus we isolated was named JX14T2. for PRRSV,. The growth characteristics of JX14T2 P5 virus were compared with those of the representative strain of high pathogenicity PRRSV and attenuated virus. The results showed that the cytopathic effect of JX14T2 on Marc-145 cells, the morphology of virus plaque and the dynamic of virus proliferation were similar to those of HP-PRRSV virus strain. The full-length gene sequencing and sequence comparison analysis of JX14T2 showed that the full-length 14960 bp, of JX14T2 was a North American virus strain. In addition to the deletion of 129 amino acids in NSP2, there were 120 amino acid deletions in 598 aa. The relationship with HP-PRRSV was close. The results of this study provide a scientific basis for mastering the prevalence and pathogenesis of PRRSV.
【作者单位】：中国农业科学院上海兽医研究所;
【基金】：973计划(2014CB542702) 国家科技支撑计划课题(2015BAD12B01-1) 国家生猪现代产业技术体系项目(CARS-36 ) 国家自然科学基金(31302098、31300140、31502072) 上海市闵行区人才发展专项资金
【分类号】：S852.651

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