猪伪狂犬病毒gB、gC和gD蛋白多抗制备及交叉中和抗体效价测定

Preparation of polyclonal antibodies against gB, gC and gD proteins of pseudorabies virus and determination of cross neutralization antibodies titers

为研究猪伪狂犬病毒（PRv）主要免疫原性基因的抗原变异情况，本研究构建了PRV变异株JS-2012和弱毒疫苗株Bartha—K61的gB、gc和gD基因重组真核表达质粒。通过基因枪途径免疫新西兰大白兔，对制备的兔源多克隆抗体进行交叉中和抗体效价的i／n,4定。结果显示，JS．2012和Bartha．K61株的gC蛋白和gD蛋白多抗对PRVJS．2012变异株、SC经典强毒株和Bartha．K61弱毒疫苗株的交叉中和效价均无显著差异；而两个病毒株的2B蛋白多抗对JS．2012和sc株的中和效价较低，分别为1：29．0～56．7和1：56．2～137．0，对Bartha—K61株以及其他毒力基因缺失弱毒疫苗株的中和效价较高，分别为1：75．0--490．0和1：198．6～986．9，差异显著，推测该结果可能与PRV毒力基因缺失有关。本研究为进一步鉴定PRV流行株的抗原变异奠定基础。

To identify the antigenic genetic variations of pseudorabies virus （PRV） main immunogenic genes, the gB, gC and gD genes from PRV JS-2012 mutant virulent strain and Bartha-K61 vaccine strain were amplified and cloned into the pcDNA3.1 to construct recombinant eukaryotic expression plasmids of pcDNA-gB, -gC and -gD, which were immunized in rabbits by gene gun for the preparation of polyclonal antibodies, respectively. The neutralization titers of antibodies were examined by cross neutralization test. There were no significant differences in the titers of gC and gD antiserum against JS-2012 and Bartha-K61 strains neutralized the JS-2012, SC （classic virulent） and Bartha-K61. However, the titers of the gB-specific antiserum against the JS-2012 and Bartha-K61 strains neutralized the JS-2012 and SC strain was significantly lower than that of Bartha-K61 and others vaccine strains. The results might be related to the genes deletions of PRV strains. This study established foundation for study genetic variation of the PRV prevalent strains.