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为了确诊猪伪狂犬病病毒(PRV)的感染,探讨目前广西猪群中流行的PRVgE基因的变异特征,为更好地防控猪伪狂犬病(PR)提供参考依据,本研究采集了广西陆川某猪场保育猪群发生呼吸道症状的肺脏组织,并用Vero细胞进行病毒分离,应用PCR方法对分离株的gE基因进行克隆和测序,根据测序结果证实分离到1个PRV毒株,命名为GXLC1。分离株在Vero细胞上增殖,细胞出现典型的病变;GXLC1株gE基因与GenBank上20株国内外具有代表性参考毒株的核苷酸序列同源性为97.1%~99.4%,氨基酸同源性为94.3%~99.6%;gE基因的遗传进化分析显示,GXLC1株与2012年的国内流行毒株亲缘关系较近,与欧美分离株亲缘关系较远;氨基酸序列分析显示,GXLC1株gE蛋白主要抗原表位区较之国内经典强毒株有3个氨基酸位点的变异,可能导致gE抗原性发生改变。本研究将分离到的1个PRV毒株进行了gE基因的分析,发现GXLC1株为近年来流行的变异株,gE蛋白在抗原表位区上有3个氨基酸位点的变异,这是否会影响GXLC1株毒力和抗原性的变化,还有待进一步研究;对gE基因变异特征的分析和病毒的分离为进一步丰富广西PRV的分子流行病学和疫苗的研制提供参考依据和重要材料。
In order to confirm the PRV infection and to explore the variation characteristics of the PRVgE gene currently prevailing in Guangxi swine herds and to provide a reference for better prevention and control of PRV, The lung tissues of the pigs with respiratory symptoms were preserved in the field. Vero cells were used for virus isolation. The gE gene of the isolate was cloned and sequenced by PCR. According to the sequencing results, one PRV strain was isolated and named as GXLC1. The isolates proliferated on Vero cells and showed typical cell lesions. The nucleotide sequence of gE gene of GXLC1 strain was 97.1% ~ 99.4% with reference strains from 20 domestic and foreign GenBanks. The amino acid homology Was 94.3% ~ 99.6%. The genetic evolution analysis of gE gene showed that GXLC1 strain had a close genetic relationship with the domestic epidemic strains in 2012 and was distantly related to the European and American isolates. The amino acid sequence analysis showed that GXLC1 mainly expressed gE protein Epitope region than the domestic classic virulent strains have 3 amino acid site variation, may lead to gE antigenic changes. In this study, one isolated strain of PRV was analyzed for gE gene. It was found that GXLC1 strain is a popular variant in recent years. The gE protein has 3 amino acid variations in the epitope region, which will affect whether GXLC1 strain virulence and antigenicity changes, remains to be further studied; gE gene mutation characteristics and virus isolation for further enrichment Guangxi PRV molecular epidemiology and vaccine development provide a reference and important material.