摘要
为了解福建省猪伪狂犬病病毒(PRV)野毒株的病原学特性及分子遗传变异情况,本研究用羊胚胎鼻甲细胞(OFTu)从疑似暴发伪狂犬病猪场死亡的育肥猪中分离到1株病毒,该病毒能够产生典型的PRV细胞病变,病毒液接种小鼠(0.1 mL/只)及成年兔(0.5 mL/只)在48~60 h死亡,均表现出典型的伪狂犬病症状。经PCR试验、电镜观察及特异性血清间接免疫荧光试验鉴定为伪狂犬病病毒,命名为MQ18株;并对其进行病毒效价的测定及gC、gE基因核苷酸和氨基酸序列差异性分析。结果显示,分离株TCID50为10-7.71/0.1 mL;将其gC和gE基因序列与国内外17个PRV参考毒株进行同源性比较,核苷酸序列同源性分别为93.5%~99.9%和97.8%~100%,氨基酸序列同源性分别为88.7%~99.8%和95.7%~100%;分子遗传进化树分析显示,gC和gE基因均与国内近6年分离的PRV变异株集聚在一起,属于同一个进化分支;与经典株相比,gC基因编码的氨基酸序列在第63~69位有1个AAASTPA的连续7个氨基酸插入,gE基因编码的氨基酸序列分别在第48位和第496位有1个天冬氨酸(D)的插入,与PRV变异株变异位点相一致。以上结果证实,从发病猪群中分离到1株PRV变异株,本研究结果为选择合适疫苗来防控猪伪狂犬病以及病原学研究提供了参考。
In order to understand the pathogenic characteristics and molecular genetics of wild porcine pseudorabies virus strains in Fu Jian province,a virus strain was isolated from suspicious PRV samples collected from dead fattening pigs using ovine fetal turbinate(OFTu)cells in 2018.Typical PRV cytopathic changes were observed and the inoculated mice(0.1 mL/animal)and adult rabbits(0.5 mL/animal)were died with typical PRV clinical symptoms within 48-60 h.The virus was identified as PRV by PCR,electron microscopy and indirect immunofluorescence technology,named as MQ18 strain.Then,the virus titer was determined and the genetic analyses were carried out with some other PRV sequences based on amino acid sequences and nucleotide sequences of the gC and gE genes.The results showed that TCID50 of the isolation was 10-7.71/0.1 mL.Compared with another 17 PRV reference sequences at home and abroad,the nucleotide sequence homology was 93.5%to 99.9%and 97.8%to 100%,and the amino acid homology was88.7%to 99.8%and 95.7%to 100%,respectively.Phylogenetic tree analyses of gC and gE amino acid sequences showed the strain in this study clustered together with another home PRV variants isolated from the last 6 years with a relatively independent branch.Compared with classical PRV strains,there was a continuous 7 amino acid insertion with AAASTPA at the sides from 63 to 69 in gC amino acid sequences.And there was an amino acid insertion(amino acid D)at the side 48 and 496 in gE gene respectively.The above results confirmed that the case was caused by the infection of a variant strain of PRV,which provides data for the selection of specific vaccine to control the disease and provides a reference for the etiology study on it.
作者
曾显成
池雪林
潘启东
陈珍珍
林琳
修金生
江斌
ZENG Xian-cheng;CHI Xue-lin;PAN Qi-dong;CHEN Zhen-zhen;LIN Lin;XIU Jin-sheng;JIANG Bin(College of Animal Science,Fujian Agricultural and Forestry University,Fuzhou 350002,China;Institute of Animal Science&Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou 350 013,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2019年第8期1025-1033,共9页
Chinese Veterinary Science
基金
福建农林大学科技创新专项基金项目(KFA17221A,KFA17222A)
国家自然科学基金项目(31602046)
关键词
伪狂犬病病毒
变异
分离鉴定
序列分析
pseudorabies virus
variation
isolation and identification
sequence analysis
