陕西省猪瘟流行毒株E0和E2全基因分子特征分析

【目的】分析目前陕西省猪瘟病毒（CSFV）流行毒株的E0、E2全基因分子特征，为猪瘟防控提供参考。【方法】根据CSFV Shimen株及HCLV株全基因序列，设计并合成了4对引物，采用RT-PCR方法，从采自陕西不同地区的猪瘟病料中扩增E0、E2全基因并进行序列测定分析。【结果】从采集的病料中成功扩增了5株猪瘟流行毒株的E0、E2全基因，这5株流行毒株E0基因核苷酸同源性在94.4％～99.8％，与参考毒株ALD、Alfort187、Brescia、CAP、Glentorf、GPE、GXWZ02、HCLV、Paderborn、Rimes和Shimen的核苷酸同源性在80.4％～96.3％，氨基酸同源性在86.5％～99.3％。5株流行毒株E2基因核苷酸同源性在91.1％～98.0％，与参考毒株的核苷酸同源性在77.7％～94.6％，氨基酸同源性在85.2％～95.9％。5株流行毒株E0 Rnase活性区域氨基酸基序位点没有变异，但导致流行毒株发生免疫逃逸的E2蛋白关键位点中有3个发生了变异。【结论】测定的陕西省猪瘟流行毒株E0、E2基因序列变异明显，其中E2蛋白关键位点变异较大。

Molecule character analysis of gene E0 and E2 of virulent classical swine fever virus isolates in Shaanxi province

【Objective】 The analysis on gene E0 and E2 molecular character of the virulent strain of classical swine fever in Shaanxi province was done to provide a reference for the prevention and control of classical swine fever.【Method】 According to Shimen and HCLV strains gene sequence,we designed and synthesized four pairs of primers using RT-PCR method,amplified and analyzed the gene sequence of gene E0 and E2 from the collection of disease tissue.【Result】 The results showed that the five strains gene E0 nucleotide homology was between 94.4%-99.8%.Compared with reference strains of ALD,Alfort187,Brescia,CAP,Glentorf,GPE,GXWZ02,HCLV,Paderborn,Rimes and Shimen,nucleotide homology was between 80.4%-96.3%,amino acid homology 86.5%-99.3%,genes E2 nucleotide homology 91.1%-98.0%,and compared with the reference strain nucleotide homology was between 77.7%-94.6%,and amino acid homology 85.2%-95.9%.E0 Rnase active site motifs were not mutated,and E2 protein had three key sites of mutation leading to immune escape of the virulent strain.【Conclusion】 Gene E0 and E2 sequence of virulent strains of Shaanxi province variation was significant,especially in key sites of E2 larger variation.