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为了解辽宁猪流行性腹泻病毒(PEDV)来源及变异特性,经PCR诊断,细胞分离,细胞病变观察、RT-PCR以及S基因序列鉴定,成功分离1株PEDV毒株(LN-2015-1)。S基因序列分析显示,与CV777毒株相比,除点突变外,分离株还出现最长9bp的插入,6bp的缺失和13bp连续突变。氨基酸也存在最长3aa插入和2aa缺失,同时存在4处连续3个及以上的氨基酸突变,抗原表位分析表明在5个表位区共发生16个氨基酸的突变。同源性分析显示与HB-HA2015毒株的同源关系最近,序列相似性为99.2%,与2010年以来国内外代表变异毒株同源性较高,达到98.5%~98.8%,而与2010年以前的传统同源性较低,在93.2%~95.6%之间;进化树分析显示该毒株与近年来国内外流行的变异毒株属于同一群,并且与HB-HA2015形成一小的分支,而与传统毒株进化距离较远。
To investigate the origin and variation of porcine epidemic diarrhea virus (PEDV) in Liaoning, one PEDV strain (LN-2015-1) was successfully isolated by PCR diagnosis, cell isolation, cytopathic effect observation, RT-PCR and S gene sequence identification. . S gene sequence analysis showed that in addition to the point mutation, compared with the CV777 strain, the isolate also showed a maximum of 9bp insertion, 6bp deletion and 13bp continuous mutation. The amino acids also had the longest 3aa insertion and the deletion of 2aa. At the same time, there were four consecutive amino acid mutations of three or more amino acids. The epitope analysis showed that a total of 16 amino acid mutations occurred in the five epitopes. Homology analysis showed that the homology with the strain HB-HA2015 was the closest, and the sequence similarity was 99.2%, which was highly homologous to the representative strains of variation at home and abroad since 2010, reaching 98.5% ~ 98.8% The traditional homology was lower from 93.2% to 95.6% years ago. Phylogenetic tree analysis showed that the strain belonged to the same group with the most popular mutants at home and abroad in recent years, and formed a small branch with HB-HA2015 , While the evolutionary distance with traditional strains far.