为了解当前湖南省猪伪狂犬病病毒(PRV)病原特性及基因变异情况,研究于2017—2018年从湖南省长沙、株洲、岳阳和怀化等4个地区疑似发生猪伪狂犬病(PR)的8个猪场采集组织病料8份,经荧光定量PCR检测,其中5份为PRV阳性,将PRV阳性组织样品处理后接种于猪肾上皮细胞(PK15)并连续传代,最终成功分离到5株病毒,且进一步PCR鉴定证明分离的5株病毒均为PRV.其后对5株PRV TCID50进行测定,同时对所有毒株的毒力或免疫相关基因(gB、gG、gH、gI、gL、gM、gE、TK和PK)进行扩增、测序及分析.根据地区来源将5株PRV分别命名为HuN-HH株、HuN-YY株、HuN-NX株、HuN-ZZ株和HuN-LY株,滴度分别为10-6.37、10-7.12、10-7.44、10-7.62和10-7.94 TCID50/100μL.进一步序列分析结果发现研究分离的5株PRV的毒力或免疫相关基因核苷酸序列相对保守,与国内2012年以后流行的PRV变异株同源性最高,与国内流行的传统株和国外疫苗株同源性相对较低,值得注意的是,HuN-HH株gG基因有一段连续的氨基酸序列插入.以上研究结果表明,研究分离的PRV毒株均属于变异株,且部分毒株特定基因序列存在较大变异,但其影响还需要进一步探究.

Serological investigation was carried out to understand of the immunization and infection status of porcine pseudorabies virus (PRV)in scaled pig farms of Hunan province.4 288 serum samples were collected in 208 different scale pig farms in 2018.ELISA method was used to detect PRV antibody in serum samp1es.The results showed that: the positive rate of PRV gB antibodies in immunized pigs was 78.1%，the positive rate of PRV gE antibodies was 21.0%，and the positive rate of pig farms was 38.9%;In different scale farms，large farms with more than 1 000 stocks，the positive rate of PRV gE antibodies was the highest，which was 22.9%.In different quarters，the positive rate of PRV gE antibodies in summer was the highest，which was 25.1%.in different herds，the positive rate of PRV gE antibodies was the highest，in sows which was 36.2%.It indicated that porcine pseudorabies is still widely prevalennt in Hunan，and wild-type virus infection is more common.