羊口疮病毒的分离鉴定及F1L基因的遗传变异分析
|
摘要
采用羊胚胎鼻甲(OFTu)细胞对湖北某山羊养殖场疑似羊口疮(Orf)的病羊唇部痂皮组织进行病毒分离,通过病理组织学、电镜形态观察及PCR试验等方法对分离的病毒进行鉴定,证实分离毒株为羊口疮病毒(Orf virus,ORFV),命名为HuB13。分析HuB13 F1L基因序列,结果F1L基因全长为1 017 bp,编码339个氨基酸,G+C含量为64.70%。遗传进化树显示,HuB13与8株山羊和6株绵羊来源的ORFV同源性分别为97.50%～99.41%和97.15%～97.35%,与牛痘病毒(PCPV)和牛丘疹性皮炎病毒(BPSV)同源性分别为88.0%～88.69%和75.15%。Bioedit软件分析发现宿主为山羊和绵羊的ORFV存在1个高变异区(148～171 bp)和6个特异性位点G186C、T187G、A541G、A765G、G856A和G982A。研究结果为开发宿主特异的Orf疫苗及致病机理研究提供参考依据。
An outbreak of contagious ecthyma(Orf) in goats in Hubei province was investigated.The virus was isolated from the scabs of infected goats using Ovis embryo turbinate(OFTu) cells.The etiology of the disease was identi?ed as Orf virus by histopathology,transmission electron microscopy and polymerase chain reaction(PCR),and was named HuB13.Analysis of F1 L sequences showed the total length was 1 017 bp,encoding 339 amino acids,and the content of G+C was 64.70%.Phylogenetic analysis of F1 L showed HuB13 shared 97.50%-99.41% with 8 ORFVs originating from goats and 97.15%-97.35% with 6 ORFVs originating from sheep,and shared 88.0%-88.69% and 75.15% with 2 pseudocowpox virus(PCPV) strains and a bovine papular stomatitis virus(BPSV) strain,respectively.Bioedit software analysis showed ORFVs originating from goats and from sheep had a high variation region(148-171 bp) and six specific sites as G186 C,T187 G,A541 G,A765 G,G856 A,G982 A.The results provided a theoretical basis for the development of the specific orf vaccine and pathological mechanism study.
An outbreak of contagious ecthyma(Orf) in goats in Hubei province was investigated.The virus was isolated from the scabs of infected goats using Ovis embryo turbinate(OFTu) cells.The etiology of the disease was identi?ed as Orf virus by histopathology,transmission electron microscopy and polymerase chain reaction(PCR),and was named HuB13.Analysis of F1 L sequences showed the total length was 1 017 bp,encoding 339 amino acids,and the content of G+C was 64.70%.Phylogenetic analysis of F1 L showed HuB13 shared 97.50%-99.41% with 8 ORFVs originating from goats and 97.15%-97.35% with 6 ORFVs originating from sheep,and shared 88.0%-88.69% and 75.15% with 2 pseudocowpox virus(PCPV) strains and a bovine papular stomatitis virus(BPSV) strain,respectively.Bioedit software analysis showed ORFVs originating from goats and from sheep had a high variation region(148-171 bp) and six specific sites as G186 C,T187 G,A541 G,A765 G,G856 A,G982 A.The results provided a theoretical basis for the development of the specific orf vaccine and pathological mechanism study.
引文
[1] 王光祥,尚佑军,王艳华,等.羊传染性脓疱病毒耐热冻干保护剂筛选及冻干曲线优化[J].中国兽医学报,2018,38(2):266-271.
[2] 张頔,赵魁,周艳龙,等.免疫共沉淀联合质谱技术筛选宿主因子CypB的互作蛋白[J].中国兽医学报,2017,37(5):905-908.
[3] DEMIRASLAN H,DINC G,DOGANAY M.An overview of Orf virus infection in humans and animals[J].Recent Pat Antiinfect Drug Discov,2017,12(1):21-30.
[4] BOGOCH I I,GANDHI R T,BIBI Y,et al.Human Orf virus infection from household exposures-United States,2009-2011[J].Mmwr Morb Mortal Wkly Rep,2012,308(14):245-248.
[5] YU Y Z,TAN Q,ZHAO W B,et al.Characterization of an Orf virus isolate from an outbreak in Heilongjiang province,China[J].Arch Virol,2017,162(10):1-7.
[6] 迟源,王好,钱爱东.羊传染性脓疱病毒的分离鉴定[J].中国预防兽医学报,2010,11(9):724-726.
[7] MAZUR C,FERREIRA I I,RANGEL FILHO F B,et al.Molecular characterization of Brazilian isolates of Orf virus[J].Vet Microbiol,2000,73(4):253-259.
[8] CHI X L,ZENG X C,LUO S H.Diagnosis and phylogenetic analysis of a multifocal cutaneous Orf virus with mixed bacterial infection outbreak in goats in Fujian province,China[J].Arch Virol,2017,162(10):1-10.
[9] YANG H B,MENG Q L,QIAO J,et al.Detection of genetic variations in Orf virus isolates epidemic in Xinjiang China[J].J Basic Microbiol,2014,54(11):1273-1278.
[10] ZHAO K,SONG D G,HE W Q,et al.Identification and phylogenetic analysis of an Orf virus isolated from an outbreak in sheep in the Jilin province of China[J].Vet Microbiol,2010,142(3/4):408-415.
[11] ZHANG K S,LIU Y J,KONG H J,et al.Comparison and phylogenetic analysis based on the B2L gene of Orf virus from goats and sheep in China during 2009-2011[J].Arch Virol,2014,159(6):1475-1479.
[12] WANG Y,YIN D D,YANG K K,et al.Isolation and identification of an ORF virus strain and phylogenetic analysis[J].J Yangzhou Univ,2017,38(2):10-14.
[13] WANG Q X,ZHU X R,YI C G,et al.Isolation and identification of Orf virus from jiangsu province and phylogenetic analysis of their B2L gene[J].Chin Ani Husbandry Vet Med,2017,44(4):950-958.
[14] CHI X L,ZENG X C,HAO W B,et al.Heterogeneity among Orf virus isolates from goats in Fujian Province,Southern China[J].PloS One,2013,8(10):e66958.
[15] BUDDLE B M,DELLERS R W,SCHURIG G G.Heterogeneity of contagious ecthyma virus isolates[J].Am J Vet Res,1984,45(1):75.
[16] CZERNY C P,WALDMANN R,SCHEUBECK T.Identification of three distinct antigenic sites in parapoxviruses[J].Arch Virol,1997,142(4):807-821.
[17] 任成林,张煜,崔红玉.快速动物组织块染色石蜡包埋制片法[J].中国兽医杂志,2004,40(6):26-27.
[18] 汪园园,童双,李威,等.羊胚胎鼻甲细胞的原代培养及其在羊口疮病毒研究中的应用[J].中国兽医科学,2013(5):470-475.
[19] DIEL D G,LUO S,DELHON G,et al.Orf virus ORFV121 encodes a novel inhibitor of NF-κB that contributes to virus virulence[J].J Virol,2011,85(5):2037.
[20] LI W,NING Z,HAO W,et al.Isolation and phylogenetic analysis of Orf virus from the sheep herd outbreak in northeast China[J].BMC Vet Res,2012,8(1):229.
[21] ZHANG K,LU Z,SHANG Y,et al.Diagnosis and phylogenetic analysis of Orf virus from goats in China:a case report[J].Virol J,2010,7(1):1-5.
[22] CHEN H Q,LI W,KUANG Z Z,et al.The whole genomic analysis of Orf virus strain HN3/12 isolated from Henan province,central China[J].BMC Vet Res,2017,13(1):260.
[23] ZHANG K Z,XIAO Y H,YU M,et al.Phylogenetic analysis of three Orf virus strains isolated from different districts in Shandong province,East China[J].J Vet Med Sci,2015,77(12):1639-1645.
[24] SCAGLIARINI A,GALLINA L,DAL P F,et al.Heparin binding activity of orf virus F1L protein[J].Virus Res,2004,105(2):107.
[25] HOSAMANI M,SCAGLIARINI A,BHANUPRAKASH V,et al.Orf:an update on current research and future perspectives[J].Expert Rev Anti Infect Ther,2009,7(7):879-893.
[2] 张頔,赵魁,周艳龙,等.免疫共沉淀联合质谱技术筛选宿主因子CypB的互作蛋白[J].中国兽医学报,2017,37(5):905-908.
[3] DEMIRASLAN H,DINC G,DOGANAY M.An overview of Orf virus infection in humans and animals[J].Recent Pat Antiinfect Drug Discov,2017,12(1):21-30.
[4] BOGOCH I I,GANDHI R T,BIBI Y,et al.Human Orf virus infection from household exposures-United States,2009-2011[J].Mmwr Morb Mortal Wkly Rep,2012,308(14):245-248.
[5] YU Y Z,TAN Q,ZHAO W B,et al.Characterization of an Orf virus isolate from an outbreak in Heilongjiang province,China[J].Arch Virol,2017,162(10):1-7.
[6] 迟源,王好,钱爱东.羊传染性脓疱病毒的分离鉴定[J].中国预防兽医学报,2010,11(9):724-726.
[7] MAZUR C,FERREIRA I I,RANGEL FILHO F B,et al.Molecular characterization of Brazilian isolates of Orf virus[J].Vet Microbiol,2000,73(4):253-259.
[8] CHI X L,ZENG X C,LUO S H.Diagnosis and phylogenetic analysis of a multifocal cutaneous Orf virus with mixed bacterial infection outbreak in goats in Fujian province,China[J].Arch Virol,2017,162(10):1-10.
[9] YANG H B,MENG Q L,QIAO J,et al.Detection of genetic variations in Orf virus isolates epidemic in Xinjiang China[J].J Basic Microbiol,2014,54(11):1273-1278.
[10] ZHAO K,SONG D G,HE W Q,et al.Identification and phylogenetic analysis of an Orf virus isolated from an outbreak in sheep in the Jilin province of China[J].Vet Microbiol,2010,142(3/4):408-415.
[11] ZHANG K S,LIU Y J,KONG H J,et al.Comparison and phylogenetic analysis based on the B2L gene of Orf virus from goats and sheep in China during 2009-2011[J].Arch Virol,2014,159(6):1475-1479.
[12] WANG Y,YIN D D,YANG K K,et al.Isolation and identification of an ORF virus strain and phylogenetic analysis[J].J Yangzhou Univ,2017,38(2):10-14.
[13] WANG Q X,ZHU X R,YI C G,et al.Isolation and identification of Orf virus from jiangsu province and phylogenetic analysis of their B2L gene[J].Chin Ani Husbandry Vet Med,2017,44(4):950-958.
[14] CHI X L,ZENG X C,HAO W B,et al.Heterogeneity among Orf virus isolates from goats in Fujian Province,Southern China[J].PloS One,2013,8(10):e66958.
[15] BUDDLE B M,DELLERS R W,SCHURIG G G.Heterogeneity of contagious ecthyma virus isolates[J].Am J Vet Res,1984,45(1):75.
[16] CZERNY C P,WALDMANN R,SCHEUBECK T.Identification of three distinct antigenic sites in parapoxviruses[J].Arch Virol,1997,142(4):807-821.
[17] 任成林,张煜,崔红玉.快速动物组织块染色石蜡包埋制片法[J].中国兽医杂志,2004,40(6):26-27.
[18] 汪园园,童双,李威,等.羊胚胎鼻甲细胞的原代培养及其在羊口疮病毒研究中的应用[J].中国兽医科学,2013(5):470-475.
[19] DIEL D G,LUO S,DELHON G,et al.Orf virus ORFV121 encodes a novel inhibitor of NF-κB that contributes to virus virulence[J].J Virol,2011,85(5):2037.
[20] LI W,NING Z,HAO W,et al.Isolation and phylogenetic analysis of Orf virus from the sheep herd outbreak in northeast China[J].BMC Vet Res,2012,8(1):229.
[21] ZHANG K,LU Z,SHANG Y,et al.Diagnosis and phylogenetic analysis of Orf virus from goats in China:a case report[J].Virol J,2010,7(1):1-5.
[22] CHEN H Q,LI W,KUANG Z Z,et al.The whole genomic analysis of Orf virus strain HN3/12 isolated from Henan province,central China[J].BMC Vet Res,2017,13(1):260.
[23] ZHANG K Z,XIAO Y H,YU M,et al.Phylogenetic analysis of three Orf virus strains isolated from different districts in Shandong province,East China[J].J Vet Med Sci,2015,77(12):1639-1645.
[24] SCAGLIARINI A,GALLINA L,DAL P F,et al.Heparin binding activity of orf virus F1L protein[J].Virus Res,2004,105(2):107.
[25] HOSAMANI M,SCAGLIARINI A,BHANUPRAKASH V,et al.Orf:an update on current research and future perspectives[J].Expert Rev Anti Infect Ther,2009,7(7):879-893.