Ge Yufang, Jiang Feiyang, Wang Sibei, Wu Heqiong, Liu Yuan, Wang Bin, Hou Wei, Yu Xiuju, Wang Haidong
College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong 030801, China.
Single Molecule Nanometry Laboratory (Sinmolab), Nanjing Agricultural University, Nanjing 210095, China.
Animals (Basel). 2023 May 26;13(11):1766. doi: 10.3390/ani13111766.
The porcine epidemic diarrhea virus (PEDV) can cause severe piglet diarrhea or death in some herds. Genetic recombination and mutation facilitate the continuous evolution of the virus (PEDV), posing a great challenge for the prevention and control of porcine epidemic diarrhea (PED). Disease materials of piglets with PEDV vaccination failure in some areas of Shanxi, Henan and Hebei provinces of China were collected and examined to understand the prevalence and evolutionary characteristics of PEDV in these areas. Forty-seven suspicious disease materials from different litters on different farms were tested by multiplex PCR and screened by hematoxylin-eosin staining and immunohistochemistry. PEDV showed a positivity rate of 42.6%, infecting the small and large intestine and mesenteric lymph node tissues. The isolated strains infected Vero, PK-15 and Marc-145 multihost cells and exhibited low viral titers in all three cell types, as indicated by their growth kinetic curves. Possible putative recombination events in the isolates were identified by RDP4.0 software. Sequencing and phylogenetic analysis showed that compared with the classical vaccine strain, PEDV SX6 contains new insertion and mutations in the S region and belongs to genotype GIIa. Meanwhile, ORF3 has the complete amino acid sequence with aa80 mutated wild strains, compared to vaccine strains CV777, AJ1102, AJ1102-R and LW/L. These results will contribute to the development of new PEDV vaccines based on prevalent wild strains for the prevention and control of PED in China.
猪流行性腹泻病毒(PEDV)可在部分猪群中引起严重的仔猪腹泻或死亡。基因重组和突变促使该病毒(PEDV)不断进化,给猪流行性腹泻(PED)的防控带来巨大挑战。收集并检测了中国山西、河南和河北部分地区接种PEDV疫苗失败仔猪的病料,以了解这些地区PEDV的流行情况和进化特征。采用多重PCR对来自不同猪场不同窝次的47份可疑病料进行检测,并通过苏木精-伊红染色和免疫组化进行筛选。PEDV阳性率为42.6%,感染小肠、大肠和肠系膜淋巴结组织。分离毒株可感染Vero、PK-15和Marc-145多种宿主细胞,其生长动力学曲线表明在这三种细胞类型中病毒滴度均较低。利用RDP4.0软件鉴定分离株中可能存在的推定重组事件。测序和系统发育分析表明,与经典疫苗株相比,PEDV SX6在S区域存在新的插入和突变,属于GIIa基因型。同时,与疫苗株CV777、AJ1102、AJ1102-R和LW/L相比,ORF3具有完整的氨基酸序列,且aa80位点发生了突变。这些结果将有助于开发基于流行野毒株的新型PEDV疫苗,用于中国PED的防控。