PEDV变异株的分离鉴定和S1基因序列分析  

Isolation and Identification of Porcine Epidemic Diarrhea Virus(PEDV)Variants and Analysis of S1 Gene

作　　者：张江[1] 马晶晶 高俊锋 韩相敏 吴汉宇 李凯亮 奉中花 赖志 ZHANG Jiang;MA Jing-jing;GAO Jun-feng;HAN Xiang-min;WU Han-yu;LI Kai-liang;FENG Zhong-hua;LAI Zhi(Shanghai Vocational College of Agriculture and Forestry,Shanghai 201699,China;Shanghai Chuanghong Biotech Co.Lid.,Shanghai 201619,China)

机构地区：[1]上海农林职业技术学院,上海松江201699 [2]上海创宏生物科技有限公司,上海松江201619

出　　处：《中国兽医杂志》2022年第6期40-44,共5页Chinese Journal of Veterinary Medicine

摘　　要：为了获得猪流行性腹泻病毒(PEDV)分离株,并对其S1基因进行序列分析,本试验从江苏2个猪场发病死亡仔猪的肠道组织分离病毒,接种至Vero细胞培养;通过实时荧光定量PCR方法鉴定PEDV、反转录PCR(RT-PCR)方法对PEDV S1基因进行扩增测序;并将10^(7.0)TCID_(50)/mL的分离毒株口服接种5头哺乳仔猪(2 mL/头),观察临床症状,统计发病率和死亡率。结果显示,经鉴定获得的2株分离株均为PEDV,分别命名为DJCY株和DJYJ株。分离毒株能在Vero上产生典型细胞病变,传代至F10代时病毒滴度分别为10^(7.80)TCID_(50)/mL和10^(8.16)TCID_(50)/mL。RT-PCR鉴别诊断S1基因测序和分析显示,2株PEDV均为变异株。接种分离株的哺乳仔猪发病率为100%,接种DJCY株的死亡率为80%,接种DJYJ株的死亡率为100%。体外和体内试验证实,2株分离毒株均为PEDV变异株,且毒力较强。In order to obtain porcine epidemic diarrhea virus(PEDV)isolate and study its S1 gene,this study inoculated intestinal disease materials from two piglets farms in Jiangsu province onto Vero cells and generated PEDV strains which were identified by real-time PCR.S1 gene was amplified from the two PEDV strains by RT-PCR and sequenced.Five piglets were inoculated orally with 10^(7.0)TCID_(50)/mL PEDV(2 mL per piglet)and observed for clinical symptoms and pathological changes,and the morbidity and mortality of piglets were calculated.The results showed that two strains were PEDV positive and designated DJCY and DJYJ.Two strains replicated well in Vero cells and induced typical cytopathic effect(CPE).The infections titers of 10 th passage of DJCY and DJYJ were 10^(7.80)TCID_(50)/mL and 10^(8.16)TCID_(50)/mL,respectively.RT-PCR and sequencing showed that the two strains were PEDV variants based on S1 gene analysis.Both DJCY and DJYJ were pathogenic to piglets,with incidence rate of 100%(both DJCY and DJYJ),and death rates of 80%(DJCY)and 100%(DJYJ),respectively.Thus,in vitro and in vivo tests in this study confirmed that the two strains were PEDV variants and highly virulent.

关 键 词：猪流行性腹泻病毒 分离鉴定 S1基因 变异株 

分 类 号：S855.3[农业科学—临床兽医学]