ACRPVM  >> Vol. 4 No. 2 (April 2015)

    高致病性猪蓝耳病病毒重庆株的分离鉴定及其变异性分析
    Identification and Genetic Diversity of Porcine Reproductive and Respiratory Syndrome Virus Isolated from Chongqing

  • 全文下载: PDF(524KB) HTML   XML   PP.7-13   DOI: 10.12677/ACRPVM.2015.42002  
  • 下载量: 806  浏览量: 4,330   国家科技经费支持

作者:  

王 璐,顾 盼,周康森,王玲玲,蔡家利:重庆理工大学药学与生物工程学院,重庆;
肖 红,陈脊宇,黎晓敏:西南大学动物科技学院,重庆;
黄 倩:重庆三峡学院,重庆

关键词:
PRRSV病毒鉴定NSP2GP5病毒变异PRRSV Viral Identification NSP2 GP5 Viral Variation

摘要:

从重庆某猪场采集病死猪病料,经RT-PCR初步鉴定其病原为猪繁殖与呼吸综合征病毒后,利用反转录合成cDNA,设计NSP2和GP5基因片段的特异性引物,并对其扩增,序列测序,分析鉴定。结果表明,该分离株的NSP2基因与经典毒株VR-2332核苷酸同源性仅为66.7%,与高致病变异株SD、JXA1的核苷酸同源性高达98.8%,GP5基因在个别碱基上也存在变异,由此表明该毒株为高致病性蓝耳病变异株。同时将该病料接种于Marc-145细胞,可观察到典型的细胞病变(CPE),经Reed-Muench计算该分离株的病毒滴度为10−5.67 TCID50/0.1mL。该毒株的分离鉴定,为进行猪繁殖与呼吸综合征流行病学的调查研究提供了依据。

The visceral organs were collected from swines at a farm in ChongQing. Once the paghogen was idetified as the porcine reproductive and respiratory syndrome virus via RT-PCR identification, we used reverse transcription to make cDNA. Both NSP2 and GP5 gene were amplified by PCR, and then they were sequenced. The NSP2 sequence analysis indicated that homology between CQ and classical VR-2332 strain was 66.7%, while it was as high as 98.8% compared with HP-PRRSV vari-ation SD and JXA1 in nucleotide. And the GP5 sequence analysis indicated that there were varia-tions in individual bases. It was suggested that the CQ strain was a HP-PRRSV variation. At the same time, the patho-material was inoculated in Marc-145 cells, and then the CPE appeared. The viral titer is 10−5.67 TCID50/0.1mL by Reed-Muench. The separation of CQ strain offers basis for ep-idemiology investigations of PRRSV.

文章引用:
王璐, 肖红, 陈脊宇, 顾盼, 周康森, 王玲玲, 黄倩, 黎晓敏, 蔡家利. 高致病性猪蓝耳病病毒重庆株的分离鉴定及其变异性分析[J]. 亚洲兽医病例研究, 2015, 4(2): 7-13. http://dx.doi.org/10.12677/ACRPVM.2015.42002

参考文献

[1] Goyal, S.M. (1993) Porcine reproductive and respiratory syndrome. Journal of Veterinary Diagnostic Investigation, 5, 656-664.
[2] Zimmerman, J.J., Yoon, K.J., Wills, R.W., et al. (1997) General overview of PRRSV: A perspective from the United States. Veterinary Microbiology, 55, 187-196.
[3] Han, K., Seo, H.W., Oh, Y., et al. (2012) Effects of North American porcine reproductive and respiratory syndrome virus (PRRSV)-based modified live vaccines on preimmunized sows artificially inseminated with European PRRSV- spiked semen. Clinical and Vaccine Immunology, 19, 319-324.
[4] 郭宝清, 陈章水, 刘文兴, 等 (1996) 从疑似PRRS流产胎儿分离PRRSV的研究. 中国畜禽传染病, 2, 1-4.
[5] Bautista, E.M., Meulenberg, J.J.M., Choi, C.S. and Molitor, T.W. (1996) Structural polypeptides of the American (VR- 2332) strain of porcine reproductive and respiratory syndrome (PRRS) virus. Archives of Virology, 141, 1357-1365.
[6] Oleksiewicz, M.B., Botner, A., Toft, P., et al. (2001) Epitope mapping porcine reproductive and respiratory syndrome virus by phage display: The nsp2 fragment of the replicase polyprotein contains a cluster of B-cell epitopes. Journal of Virology, 75, 3277-3290.
[7] 刘光清, 蔡雪辉, 仇华吉, 等 (2001) 猪生殖和呼吸系统综合征的研究进展. 中国预防兽医学报, 1, 72-76.
[8] Oleksiewicz, M.B., Botner, A. and Normann, P. (2002) Porcine B-cells recognize epitopes that are conserved between the structural proteins of American and European type porcine reproductive and respiratory syndrome virus. The Journal of General Virology, 83, 1407-1418.
[9] 王旭荣, 马红艳, 祝卫国, 等 (2008) 高热病猪群中PRRSV的调查及其流行毒株的分离. 中国兽医杂志, 3, 39- 40.
[10] Ostrowski, M., Galeota, J.A., Jar, A.M., et al. (2002) Identification of neutralizing angnonneutralizing epitopes in the porcine reproductive and respiratory syndrome virus GP5 ectodomain. Journal of Virology, 76, 4241-4250.
[11] 米自由, 熊仲良, 等 (2013) 重庆地区高致病性猪蓝耳病病原变异及免疫防控技术研究. 国家科技成果.
[12] 杨泽林, 曾政, 等 (2010) 重庆市高致病性猪蓝耳病流行病学调查. 中国动物检疫, 12, 56-58.
[13] 邢海云, 梅林, 等 (2011) 猪繁殖与呼吸综合征弱毒疫苗的安全性及新型疫苗的研究进展. 中国生物制品学杂志, 2, 237-240.
[14] Nam, H.-M., Chae, K.-S., et al. (2013) Immune responses in mice vaccinated with virus-like particles composed of the GP5 and M proteins of porcine reproductive and respiratory syndrome virus. Archives of Virology, 158, 1275-1285.