摘要
为了解内蒙古自治区伪狂犬病毒(PRV)的流行变异情况,本研究从内蒙古某猪场采集疑似患猪伪狂犬病(PR)的猪脑组织,处理后采用PRV gE基因特异性引物经PCR鉴定后将阳性样品分别接种MDBK细胞和Vero细胞,分别进行病毒的分离,对分离的病毒测定病毒含量(TCID_(50))。PCR结果显示,组织样品为PRV野毒感染。病料样品接种MDBK细胞和Vero细胞盲传4代后均产生明显的细胞病变,病毒含量为108.8 TCID_(50)/mL。表明分离到1株PRV,命名为HS01株。采用PCR分别扩增分离病毒的gE、gC和TK基因并进行同源性、遗传进化及主要氨基酸位点的变异分析。结果显示,HS01株gE、gC、TK基因序列与2011年后国内变异株相应基因的同源性分别为96.9%~99.7%、97.6%~99.6%和99.7%~100%;且他们的系统进化树结果显示,PRV HS01株与国内变异株属于同一分支,而与国内经典株遗传距离较远(gE、gC基因),或与部分国内经典株属于同一分支(TK基因),与同源性分析结果基本相符;HS01株gE基因在aa48和aa492各插入一个天冬氨酸,符合PRV变异株的特点。上述结果均表明,HS01为一株PRV变异株。将该株病毒经滴鼻接种豚鼠,进行致病力试验。结果显示接种12 h后豚鼠出现食欲不振、精神沉郁、瘙痒等临床症状,并于接种后66 h出现死亡。将感染组存活豚鼠迫杀后显示肺脏出血、充血,表明HS01株对豚鼠有致病性,并能致其死亡。本研究首次在内蒙古地区猪场分离到PRV变异株,并进行了豚鼠的致病力研究,为内蒙古自治区PRV新流行株遗传变异机制的研究及新型疫苗研发提供参考。
In order to know the probability of genetic mutation for the prevalent pseudorabies viru(PRV)isolates in Inner Mongolia,the samples of brain tissues from the suspected pseudorabies virus infected pigs were collected in Inner Mongolia,virus isolation and identification were performed for the samples with gE gene-specific PCR positive results by using MDBK and Vero cells,and the virulence obtaining viral isolates was determined by TCID_(50).The results showed that the tissue samples were infected with PRV wild virus.The viral isolates could cause the obviously cytopathic effect after being passaged blindly for four generations and the virus content could reach 108.8TCID_(50)/mL.It is suggesting that,a pseudorabies virus mutant was isolated and was named HS01.The gE,gC and TK genes of this isolate were amplified by PCR and the homology,genetic evolution and variation of resulting sequences were analyzed on the main amino acid sites.And the homologies of the new isolate was 96.9%-99.7%,97.6%-99.6%and 99.7%-100%,compared with different PRV stains isolated after 2011,respectively.The results of phylogenetic tree showed that the PRV HS01 strain belongs to the same branch as the mutant strain in our country,but is genetically distant from the classical strain(gE and gC genes)in our country,or belongs to the same branch with some classical strains(TK gene)in our country,which is basically consistent with the results of homology analysis;HS01 strain gE gene was inserted by an aspartic acid at both aa48 and aa492 sites,which is in line with the characteristics of PRV mutant strains.All the above results show that HS01 is a PRV mutant strain.This strain was inoculated with guinea pigs through nose drops,and the pathogenicity test was carried out.The results showed that the guinea pigs developed clinical symptoms such as loss of appetite,depression,and itching after 12 hours vaccination,and died at 66 hours.The surviving guinea pigs in the infected group showed lung hemorrhage and congestion after forced killing,indicating th
作者
昝晓慧
孙瑶
王岩
马艳华
柴春霞
付存
王世荣
王镓磊
武有志
阴花
王炜
ZAN Xiao-hui;SUN Yao;WANG Yan;MAYan-hua;CHAI Chun-xia;FU Cun;WANG Shi-rong;WANG Jia-lei;WU You-zhi;YIN Hua;WANG Wei(State Key Laboratory of Reproductive Regulation&breeding of grassland livestock,Inner Mongolia University,Hohhot 010070,China;Nanjing Agricultural University,Nanjing 210095,China;China Animal Husbandry Industry Co.,Ltd.,Beijing 100070,China;Basic Medical School,Inner Mongolia Medical University,Hohhot 010110,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2021年第10期1040-1045,共6页
Chinese Journal of Preventive Veterinary Medicine
基金
2020年“草原英才”(12000-12102520)。
关键词
猪伪狂犬病毒
变异株
豚鼠
致病力
基因分析
porcine pseudorabies virus
variant strain
guinea pig
pathogenicity
gene analysis
