2013—2018年广西地区猪伪狂犬病病毒gB、gE与TK基因遗传变异分析

doi:10.11843/j.issn.0366-6964.2020.04.017

摘要/Abstract

摘要： 为了解广西近年猪伪狂犬病（PR）流行情况及猪伪狂犬病病毒（PRV）流行株gB、gE、TK基因的遗传变异特点，于2013-2018年采集广西各地区疑似PR发病猪靶组织714份，进行PRV的PCR检测及病毒分离鉴定，并运用DNAStar、MEGA 6.0等生物学软件对分离毒株gB、gE、TK基因进行遗传变异分析。结果显示：样品检测阳性率为24.5%（175/714），共分离获得PRV流行株38株。受检猪群普遍存在PRV感染，经典毒株及变异毒株并存，而且以变异毒株为主。广西流行株不同亚群PRV在gB、gE、TK基因上均存在相同的氨基酸变异特点，与国内变异株亲缘关系较近。推测广西流行株存在两种不同重组变异形式产生的PRV重组毒株，分别为疫苗株gB基因和变异株gE基因重组产生的重组毒株，以及经典株的TK基因被疫苗株（HB98）的TK基因替换而产生的重组毒株。近年广西受检猪场较普遍存在PRV感染，而且PRV流行株以变异毒株为主，可能存在两种不同重组变异形式产生的重组毒株，猪场需重视及实施针对性PR免疫防控计划。

关键词: 猪伪狂犬病病毒(PRV), gB基因, gE基因, TK基因, 序列分析, 遗传变异

Abstract: To study the epidemic situation of pseudorabies (PR) and genetic variation of epidemic strains of pseudorabies virus(PRV)in Guangxi from 2013 to 2018, 714 target tissues of suspected PR-infected pigs in various regions of Guangxi were collected and tested by PCR and virus isolation. gB, gE and TK genes of isolated strains were amplified, cloned, sequenced and analyzed by DNAStar and MEGA 6.0. The results indicated that the positive rate of suspected tissues was 24.5% (175/714), and 38 PRV strains were isolated. PRV infection of Pig is common in the tested herds. There co-existed classical strains and the variant strains, and the mutant strains are dominant. gB, gE and TK genes of different subtypes of isolated strains had the same amino acid variation characteristics and had closer relationship with domestic mutant strains. It is speculated that there were two different recombinant forms of PRV strains in Guangxi. The first was recombinant by vaccine strains (gB gene) and mutant strains (gE gene). The other one, classical strain (TK gene) was replaced by vaccine strains (TK gene). In recent years, PRV infection was common in the pig farms in Guangxi, and the epidemic strains were mainly mutant strains. And there may be two PRV recombinant forms of recombinant variant strains. Pig farms should pay more attention to practice PR targeted immunity plan.

Key words: porcine pseudorabies virus (PRV), gB gene, gE gene, TK gene, sequence analysis, genetic evolution analysis

中图分类号:

S852.659.1

赵硕, 王若木, 党佳佳, 许力士, 秦树英, 薛辉, 韦祖樟, 陈樱, 欧阳康, 黄伟坚. 2013—2018年广西地区猪伪狂犬病病毒gB、gE与TK基因遗传变异分析[J]. 畜牧兽医学报, 2020, 51(4): 810-819.

ZHAO Shuo, WANG Ruomu, DANG Jiajia, XU Lishi, QIN Shuying, XUE Hui, WEI Zuzhang, CHEN Ying, OUYANG Kang, HUANG Weijian. Genetic Variation Analysis of gB, gE and TK Genes of Pseudorabies Virus in Guangxi from 2013 to 2018[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(4): 810-819.

参考文献 30

[1]	殷震. 动物病毒学[M].2版.北京:科学出版社,1997.YIN Z. Animal virology[M]. 2nd ed. Beijing:Science Press, 1997. (in Chinese)
[2]	刘涛, 王瑞, 曲哲会, 等. 2014-2016年豫南地区规模化猪场猪主要病毒性传染病流行病学调查分析[J]. 黑龙江畜牧兽医, 2018(20):106-108.LIU T, WANG R, QU Z H, et al. Epidemiological survey and analysis of major viral infectious diseases in large-scale pig farms in southern Henan from 2014 to 2016[J]. Heilongjiang Animal Science and Veterinary Medicine, 2018(20):106-108. (in Chinese)
[3]	赵雪丽, 班付国, 马震原, 等. 猪伪狂犬病免疫抗体及感染抗体消长规律研究[J]. 现代牧业, 2019, 3(1):14-16.ZHAO X L, BAN F G, MA Z Y, et al. Study on the change law of anti-pseudorabies virus immune antibody and infected antibody[J]. Modern Animal Husbandry, 2019, 3(1):14-16. (in Chinese)
[4]	张青占. 变异猪伪狂犬病毒的分离鉴定及生物学特性分析[D]. 北京:中国农业科学院, 2013.ZHANG Q Z. Isolation and identification of a mutated swine pseudorabies virus and its biological characteristics analysis[D]. Beijing:Chinese Academy of Agricultural Sciences, 2013. (in Chinese)
[5]	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):1-4.PENG J M, AN T Q, ZHAO H Y, et al. Identification and antigenic variation of new epidemiology of pseudorabies virus from swine[J]. Chinese Journal of Preventive Veterinary Medicine, 2013, 35(1):1-4. (in Chinese)
[6]	王国栋. 江山地区猪伪狂犬的流行现状与防控措施[J]. 畜禽业, 2019, 30(3):47.WANG G D. The epidemic status and control measures of pig pseudorabies in Jiangshan area[J]. Livestock and Poultry Industry, 2019, 30(3):47. (in Chinese)
[7]	李海琴, 康昭风, 谭美芳, 等. 江西省猪伪狂犬病原流行病学调查及其gB、gE及TK基因遗传变异分析[J]. 江西农业大学学报, 2019, 41(2):332-339.LI H Q, KANG Z F, TAN M F, et al. Epidemiological survey of pseudorabies in Jiangxi province and analysis of genetic variation of gB, gE and TK genes[J]. Acta Agriculturae Universitatis Jiangxiensis, 2019, 41(2):332-339. (in Chinese)
[8]	LUO Y Z, LI N, CONG X, et al. Pathogenicity and genomic characterization of a pseudorabies virus variant isolated from Bartha-K61-vaccinated swine population in China[J]. Vet Microbiol, 2014, 174(1-2):107-115.
[9]	DEAN H J, MILLER J M, ACKERMANN M R, et al. Replication and pathogenicity after intranasal and intracranial inoculation of swine with a recombinant pseudorabies virus containing a deletion at the UL/IR junction[J]. Virology, 1996, 223(1):19-28.
[10]	秦树英. 猪伪狂犬病毒GXLB-2015和GXGG-2016毒株的全基因组序列分析和生物学特性研究[D]. 南宁:广西大学, 2017.QIN S Y. Complete genome amalysis and biological characteristics of PRV GXLB-2015 and GXGG-2016 strain[D]. Nanning:Guangxi University, 2017. (in Chinese)
[11]	庞旋飞, 练斯南, 伍建敏, 等. 伪狂犬病病毒FS-2015株gE和gB基因序列分析[J]. 中国畜牧兽医, 2018, 45(12):3524-3534.PANG X F, LIAN S N, WU J M, et al. Sequence analysis of gE and gB genes of pseudorabies virus FS-2015 strain[J]. China Animal Husbandry & Veterinary Medicine, 2018, 45(12):3524-3534. (in Chinese)
[12]	LUKÀCS N, THIEL H J, METTENLEITER T C, et al. Demonstration of three major species of pseudorabies virus glycoproteins and identification of a disulfide-linked glycoprotein complex[J]. J Virol, 1985, 53(1):166-173.
[13]	LI X D, YANG F L, HU X L, et al. Two classes of protective antibodies against Pseudorabies virus variant glycoprotein B:implications for vaccine design[J]. PLoS Pathog, 2017, 13(12):e1006777.
[14]	任卫科, 池晶晶, 李秀丽, 等. 伪狂犬病毒主要毒力基因的研究进展[J]. 畜牧兽医科学, 2017(8):12-13.REN W K, CHI J J, LI X L, et al. Research progress on the main virulence genes of Pseudorabies virus[J]. Graziery Veterinary Sciences, 2017(8):12-13. (in Chinese)
[15]	李宁, 吴志明, 闫若潜, 等. 2012-2014年河南省猪伪狂犬病病毒gE基因和TK基因的遗传变异分析[J]. 中国兽医学报, 2016, 36(10):1653-1657.LI N, WU Z M, YAN R Q, et al. Genetic variation analysis of gE and TK genes of pseudorabies virus in Henan province from 2012 to 2014[J]. Chinese Journal of Veterinary Science, 2016, 36(10):1653-1657. (in Chinese)
[16]	薛辉. 猪伪狂犬病毒变异株的分离鉴定及主要基因序列分析和生物学特性研究[D]. 南宁:广西大学, 2015.XUE H. Isolation and identification, major gene sequence analysis of pseudorabies virus variant strain and its biological characteristics study[D]. Nanning:Guangxi University, 2015. (in Chinese)
[17]	周金柱. Bartha-k61株疫苗对猪伪狂犬病病毒变异株的免疫保护及变异株生物学特性的初步研究[D]. 扬州:扬州大学, 2018.ZHOU J Z. Protection efficacy of Bartha-k61 vaccine strain in growing pigs against emerging variant pseudorabies virus challenge and the preliminary biological characteristics of the variant pseudorabies viruses[D]. Yangzhou:Yangzhou University, 2018. (in Chinese)
[18]	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.
[19]	白挨泉. 新型猪伪狂犬病流行情况及净化措施[J]. 兽医导刊, 2016(3):22-24.BAI A Q. Epidemic situation and purification measures of new swine pseudorabies[J]. Veterinary Orientation, 2016(3):22-24. (in Chinese)
[20]	陈焕春, 周复春,方六荣, 等. 伪狂犬病病毒鄂A株TK-/gG-/LacZ+突变株的构建[J]. 病毒学报, 2001, 17(1):69-74.CHEN H C, ZHOU F C, FANG L R, et al. Construction of TK-/gG-/LacZ+ mutant of pseudorabies virus strain Ea[J]. Chinese Journal of Virology, 2001, 17(1):69-74. (in Chinese)
[21]	余秋颖, 常洪涛, 陈文定, 等. 2012-2013年新流行猪伪狂犬病病毒的分离鉴定及其gE、TK、gD基因序列分析[J]. 中国兽医学报, 2014, 34(10):1573-1578, 1583.YU Q Y, CHANG H T, CHEN W D, et al. Isolation and identification of pseudorabies virus prevailing from 2012 to 2013 and sequence analysis of gE, TK, gD gene[J]. Chinese Journal of Veterinary Science, 2014, 34(10):1573-1578, 1583. (in Chinese)
[22]	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.ZHOU C L, TAN X, XU T, et al. Identification and antigenic variation of variant PRV[J]. Chinese Journal of Veterinary Science, 2017, 37(3):404-409. (in Chinese)
[23]	叶超. 猪伪狂犬病病毒HeN1株全基因组测序与病毒遗传变异分析[D]. 北京:中国农业科学院, 2015.YE C. Genomic sequencing of pseudorabies virus HeN1 strain and genetic diversity analysis of the virus[D]. Beijing:Chinese Academy of Agricultural Sciences, 2015. (in Chinese)
[24]	王一鹏, 王亚文,徐瑞涛,等.一株分离自免疫猪场的伪狂犬病病毒的鉴定与变异分析[J].畜牧兽医学报,2019,50(10):2070-2078.WANG Y P, WANG Y W, XU R T, et al. Indentification and variation analysis of pseudorabies virus from vaccinated pig farm[J]. Acta Veterinaria et Zootechnica Sinica,2019,50(10):2070-2078.(in China)
[25]	王宇, 李段, 张乐宜, 等. 猪伪狂犬病毒的分离鉴定及其gE、TK全基因序列遗传变异分析[J]. 广东农业科学, 2016, 43(7):133-144.WANG Y, LI D, ZHANG L Y, et al. Isolation and identification of porcine pseudorabies virus and genetic variation analysis of gE, TK gene[J]. Guangdong Agricultural Sciences, 2016, 43(7):133-144. (in Chinese)
[26]	孙佳楠, 王楠楠, 王瑞, 等. 猪伪狂犬病病毒LN1301株分离鉴定及其gE和TK基因的变异分析[J]. 中国兽医学报, 2016, 36(8):1324-1329.SUN J N, WANG N N, WANG R, et al. Isolation and identification of pig pseudorabies virus strain LN1301 and its gE and TK gene mutation analysis[J]. Chinese Journal of Veterinary Science, 2016, 36(8):1324-1329. (in Chinese)
[27]	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]. 北京:中国农业科学院, 2014.ZHAO H Y. Identification of pseudorabies virus variants in swine and the research of their biological characteristics[D]. Beijing:Chinese Academy of Agricultural Sciences, 2014. (in Chinese)
[28]	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.GAO Z W, FAN G F, ZHAO T T, et al. Phylogenetic and sequence alignment analysis of epidemic swine Pseudorabies virus[J]. Chinese Journal of Veterinary Science, 2018, 38(1):1-10. (in Chinese)
[29]	陈超. 山东省免疫猪场猪伪狂犬病毒分离鉴定及gE、gC、gD和TK基因的序列分析[D]. 泰安:山东农业大学, 2015.CHEN C. Sequence analysis of gE, gC, gD and TK gene among pseudorabies virus strains[D]. Tai'an:Shandong Agricultural University, 2015. (in Chinese)
[30]	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.HUA L Z, LIU J F, FENG Z X, et al. Progress in novel pseudorabies virus variants prevalent in China[J]. Jiangsu Journal of Agricultural Sciences, 2017, 33(2):476-480. (in Chinese)