山东省部分地区猪伪狂犬病病毒分离鉴定及其gE基因遗传变异分析
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摘要
自2011年下半年以来全国多个省份使用Bartha-K61疫苗(gE/gI双缺失)免疫的猪场陆续出现了母猪繁殖障碍、仔猪神经症状及死亡、伪狂犬gE抗体阴性猪群突然转阳的问题。同时有的猪场内出现犬、猫、鼠等死亡的现象,经检测都是由变异伪狂犬病病毒(vPRV)引起。为了解山东省猪伪狂犬病病毒(PRV)近期流行特征,本试验对2017年上半年来自山东省不同地区的12份伪狂犬疑似病料进行了PRV的分离鉴定,并对分离株的gE基因进行了序列测定及遗传变异分析。结果表明,共分离到9株PRV,其核苷酸和氨基酸序列的同源性分别为97.5%～99.4%和97.2%～99.1%,与参考毒株其核苷酸和氨基酸序列的同源性分别为97.5%～99.8%和96.8%～99.2%,并且9株分离株与国内报道的PRV变异株的核苷酸和氨基酸序列的同源性均高于欧美毒株。gE基因的遗传进化树分析表明,本试验中9株分离株相互之间遗传距离较近,与国内PRV变异株处在同一分支,而与欧美毒株遗传距离较远。
In latter 2011,pig farms in many provinces of China vaccinated with the Bartha-K61vaccine(gE/gI double deletion)showed reproductive disorders of sows,neurologic symptoms and death of piglets,and the sudden transformation of gE antibody from negative to positive.Meanwhile,there were the death of dogs,cats,mice and other in some pig farms,which were caused by the variant pseudorabies virus(vPRV)through detection.To understand the recent epidemic characteristics of pseudorabies virus(PRV)in Shandong province,our research conducted isolation,identification and phylogenetic analysis of gE gene of pseudorabies virus on 12 samples collected from parts of Shandong province in the first half of 2017.The results showed that 9strains of PRV were isolated and they shared nucleotide and amino acid homologies of 97.5%-99.4% and 97.2%-99.1%.While nucleotide and amino acid homologies were 97.5%-99.8% and 96.8%-99.2% when compared with the reference strains,and the 9isolates shared higher nucleotide and amino acid identities with Chinese variant strains than Euramerican strains.Phylogenetic analysis of gE gene showed that the 9isolates shared close genetic distance with vPRV and were clustered into the same branch,while were distant from Euramerican strains.
In latter 2011,pig farms in many provinces of China vaccinated with the Bartha-K61vaccine(gE/gI double deletion)showed reproductive disorders of sows,neurologic symptoms and death of piglets,and the sudden transformation of gE antibody from negative to positive.Meanwhile,there were the death of dogs,cats,mice and other in some pig farms,which were caused by the variant pseudorabies virus(vPRV)through detection.To understand the recent epidemic characteristics of pseudorabies virus(PRV)in Shandong province,our research conducted isolation,identification and phylogenetic analysis of gE gene of pseudorabies virus on 12 samples collected from parts of Shandong province in the first half of 2017.The results showed that 9strains of PRV were isolated and they shared nucleotide and amino acid homologies of 97.5%-99.4% and 97.2%-99.1%.While nucleotide and amino acid homologies were 97.5%-99.8% and 96.8%-99.2% when compared with the reference strains,and the 9isolates shared higher nucleotide and amino acid identities with Chinese variant strains than Euramerican strains.Phylogenetic analysis of gE gene showed that the 9isolates shared close genetic distance with vPRV and were clustered into the same branch,while were distant from Euramerican strains.
引文
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[15]周春陵,谭鑫,许拓,等.PRV变异株的分离及抗原差异性分析[J].中国兽医学报,2017,37(3):404-409.
[16]刘曜综,芮萍,王秋悦,等.猪流行伪狂犬病gE基因检测及生物信息学分析[J].中国兽医学报,2015,35(4):536-539+544.
[17]华利忠,刘剑锋,冯志新,等.猪伪狂犬病病毒新流行变异毒株的研究进展[J].江苏农业学报,2017,33(2):476-480.
[18]赵鸿远.猪伪狂犬病病毒变异株的分离鉴定及其生物学特性的研究[D].黑龙江哈尔滨:哈尔滨兽医研究所,2014.
[19]张青占.变异猪伪狂犬病毒的分离鉴定及其生物学特性分析[D].上海:上海兽医研究所,2013.
[20]陈超,刘存,李海娥,等.山东省免疫猪场猪伪狂犬病病毒分离鉴定及gE毒力基因的序列分析[J].中国兽医学报,2016,36(6):902-907.
[21] WANG C H,YUAN J,QIN H Y,et al.A novel gEdeleted pseudorabies virus(PRV)provides rapid and complete protection from lethal challenge with the PRV variant emerging in Bartha-K61-vaccinated swine population in China[J].Vaccine,2014,32(27):3379-3385.
[2]高泽文,范桂芳,赵婷婷,等.猪伪狂犬病病毒流行株遗传进化及序列比对分析[J].中国兽医学报,2018,38(1):1-10.
[3]POMERANZ L E,REYNOLDS A E,HENGARTNER C J.Molecular biology of pseudorabies virus:impact on neurovirology and veterinary medicine[J].Microbiol Mol Biol Rev,2005,69(3):462-500.
[4]张玲玲,时建立,彭喆,等.猪伪狂犬病病毒山东分离株gE和gD基因的克隆及序列分析[J].中国兽医学报,2017,37(11):2048-2055.
[5]HAN J,CHADHA P,STARKEY J L,et al.Function of glycoprotein e of herpes simplex virus requires coordinated assembly of three tegument proteins on its cytoplasmic tail[J].Proc Natl Acad Sci USA,2012,109(48):19798-19803.
[6]刘正飞,陈焕春,吴斌,等.伪狂犬病病毒鄂A株TK-/gE-/gI-基因缺失疫苗的安全性和保护力研究[J].畜牧兽医学报,2004,35(1):70-73.
[7]METTENLEITER T C.Immunobiology of pseudorabies(Aujeszky's disease)[J].Vet Immunol Immunopathol,1996,54(1/4):221-229.
[8]SPEAR P G,EISENBERG R J,COHEN G H.Three classes of cell surface receptors for alpha-herpesvirus entry[J].Virology,2000,275(1):1-8.
[9]王勤.猪伪狂犬病病毒gE基因的研究进展[J].畜牧与兽医,2002,34(10):42-44.
[10] YU X L,ZHOU Z,HU D M,et al.Pathogenic pseudorabies virus,China,2012[J].Emerg Dis,2014,20(1):102-104.
[11] WU R,BAI C Y,SUN J Z,et al.Emergence of virulent pseudorabies virus infection in northern China[J].J Vet Sci,2013,14:363-365.
[12]童武,张青占,郑浩,等.免疫后发病仔猪中伪狂犬病毒的分离和鉴定[J].中国动物传染病学报,2013,21(3):1-7.
[13] AN T Q,PENG J M,TIAN Z J,et al.Pseudorabies virus variant in Bartha-K61-vaccinated pigs,China,2012[J].Emerg Infect Dis,2013,19(11):1749-1755.
[14]彭金美,安同庆,赵鸿远,等.猪伪狂犬病病毒新流行株的分离鉴定及抗原差异性分析[J].中国预防兽医学报,2013,35(1):1-6.
[15]周春陵,谭鑫,许拓,等.PRV变异株的分离及抗原差异性分析[J].中国兽医学报,2017,37(3):404-409.
[16]刘曜综,芮萍,王秋悦,等.猪流行伪狂犬病gE基因检测及生物信息学分析[J].中国兽医学报,2015,35(4):536-539+544.
[17]华利忠,刘剑锋,冯志新,等.猪伪狂犬病病毒新流行变异毒株的研究进展[J].江苏农业学报,2017,33(2):476-480.
[18]赵鸿远.猪伪狂犬病病毒变异株的分离鉴定及其生物学特性的研究[D].黑龙江哈尔滨:哈尔滨兽医研究所,2014.
[19]张青占.变异猪伪狂犬病毒的分离鉴定及其生物学特性分析[D].上海:上海兽医研究所,2013.
[20]陈超,刘存,李海娥,等.山东省免疫猪场猪伪狂犬病病毒分离鉴定及gE毒力基因的序列分析[J].中国兽医学报,2016,36(6):902-907.
[21] WANG C H,YUAN J,QIN H Y,et al.A novel gEdeleted pseudorabies virus(PRV)provides rapid and complete protection from lethal challenge with the PRV variant emerging in Bartha-K61-vaccinated swine population in China[J].Vaccine,2014,32(27):3379-3385.