【摘要】 为了分离猪流行性腹泻病毒(porcine epidemic diarrhea virus, PEDV)变异毒株，了解其进化趋势及致病性，并为后续PEDV特性及疫苗研发提供候选毒株，本研究将PEDV阳性样品接种于Vero细胞传代，采用间接免疫荧光试验(IFA)对分离株进行鉴定，基于RT-PCR及第2代测序技术进行全基因组测序、同源性比对及遗传进化分析，通过动物致病性试验进行毒力测定。IFA表明2株分离株能与PEDV S蛋白抗体发生特异性反应，确定为PEDV。2株毒株与参考毒株核苷酸同源性为95.6%～98.7%,属于变异毒株GⅡb群。分离的2株毒株分别命名为CH-HNKF-03和CH-HNPDS-01株。CH-HNKF-03和CH-HNPDS-01对哺乳仔猪攻毒后18 h即可发病，于24～56 h均出现典型的水样腹泻、呕吐、消瘦、脱水等症状，96 h内全部死亡。综上所述，本研究获得2株GⅡb亚群PEDV变异强毒株，为新型PEDV疫苗研发提供了候选毒株。更多还原

【Abstract】 In order to understand the genetic variation and pathogenicity for porcine epidemic diarrhea virus(PEDV) epidemic strains, PEDV were isolated from PEDV-positive samples using Vero cells, and identified by indirect immunofluorescence assay(IFA).Whole genome sequencing, homology comparison and genetic evolution analysis of the isolates were conducted based on RT-PCR and the second-generation sequencing technology.The virulence of the isolates was determined by pathogenic trial in suckling piglets.IFA results showed that the isolates reacted specifically with PEDV S protein antibody, which was identified as PEDV.The nucleotide homology between the two strains and the reference strain was 95.6%-98.7%,which belonged to the GⅡb group.The two isolates were named as CH-HNKF-03 and CH-HNDS-01 respectively.Disease onset occurred at 18 h after challenge with CH-HNKF-03 and CH-HNPDS-01 to suckling piglets, and typical watery diarrhea, vomiting, emaciation, dehydration and other symptoms appeared 24-56 h, and all died within 96 h.In conclusion, two highly virulent PEDV GⅡb variants were isolated in this study, which provides candidate strains for the development of novel PEDV vaccine.更多还原