| 摘要: |
| [目的]对2008年河南省猪呼吸与繁殖综合征病毒(PRRSV)流行株进行分离鉴定,并对其生物学特性进行研究,以了解免疫猪群发病的原因.[方法]从河南省猪场采集发病猪的血清和组织,采用Marc-145细胞对其中的PRRSV进行分离及初步鉴定.应用RT-PCR法对分离株进行扩增,并构建克隆载体,对阳性质粒测序后进行ORF5基因核苷酸及推导氨基酸序列分析,同时对分离毒株进行非免疫猪人工感染试验,确定其致病性.[结果]从发病猪组织和血清中分离到1株PRRSV(HN-08株),其可引起Marc-145细胞产生细胞病变(CPE),可在Marc-145细胞中稳定传代,能被PRRSV高免血清特异性中和;从人工感染发病猪组织和血清中均分离到PRRSV.RT-PCR得到与预期大小相符的特异片段,提交NCBI鉴定为HN-08株,属美洲型PRRSV.[结论]从河南省发病猪体内分离的HN-08株属美洲型PRRSV,对猪具有很强的致病力,可引起Marc-145细胞产生CPE,病毒滴度TCID50为105.1/mL. |
| 关键词: 猪繁殖与呼吸综合征 分离鉴定 生物学特性 |
| DOI： |
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| 基金项目:国家重点基础研究发展规划(973计划) |
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| Isolation, identification and some biological characteristics of PRRSV HN-08 strain |
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| Abstract: |
| 【Objective】 PRRSV epidemic strains of Henan Province were isolated and identified and its biological characteristics were studied in 2008 to understand the aetiological agent of immunized swine flocks.【Method】 PRRSV was isolated and preliminarily identified from tissues and serum samples of pigs collected in Henan using Marc-145 cell.The isolates were amplified by RT-PCR,and the clonal plamic were constructed and sequenced,then the nucleotide and deduced amino acid of ORF5 gene sequence were analysed.The artificial infection test of non immunization swine were carried out to determined the pathogenicity of the isolated strain.【Result】 A PRRSV strain (HN-08 strain) was isolated from tissues and serum samples of pigs in Henan province,the HN-08 strain was able to induce CPE typical of PRRSV on MARC-145 and stably propagated in vitro and it could be recognised by PRRSV hyperimmune serum.PRRSV was isolated both in tissues and serum samples of artifically infected pigs.A special gene sequence was obtained by RT-PCR,matched with the expected result.The sequence analysis showed that HN-08 strain belongs to the North American type of PRRSV.【Conclusion】 These results suggested that HN-08 strain isolated from infected pigs in Henan province belongs to the North American type of PRRSV,and the virus has a high pathogenicity to pigs,which was able to induce CPE typical of PRRSV on MARC-145 and the virus titers TCID50 was105.1/ mL. |
| Key words: porcine reproductive and respiratory syndrome PRRSV isolation and identification biological characteristics |
