猪A群轮状病毒BJ株的分离与鉴定
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摘要
轮状病毒是引起幼龄动物和儿童病毒性腹泻的主要病原,轮状病毒的分离鉴定为该病的流行病学调查和分子生物学特性研究奠定了基础。本研究采集北京某猪场腹泻发病仔猪肠内容物,将其接种MA104细胞,分离得到一株能产生明显细胞病变的病毒。对分离毒株进行胶体金、实时荧光定量RT-PCR和免疫荧光等方法鉴定,并对分离毒株的VP4、VP6和VP7基因进行测序及序列分析。结果表明,分离毒株为猪A群轮状病毒。按照A群轮状病毒的最新分类方法,确定该分离株VP4、VP6和VP7基因的基因型分别为P[13]型、I5型和G11型。因此,将该分离株命名为Rotavirus A pig/China/BJ/2015/G11P[13]。
Rotavirus infections are a major cause of viral diarrheas in young animals and children.Isolation and identification of rotavirus make a contribution to epidemiological survey and molecular biology study.A strain of porcine rotavirus was isolated in MA104 cell cultures from the intestinal contents of piglets with diarrhea in Beijing.The virus was identified to be porcine rotavirus by immunochromatography strip test,Real-time RT-PCR,immunofluorescence test and sequencing analysis.According to the sequence analysis,the virus was classified as group A porcine rotavirus,the genotype of VP4,VP6 and VP7genes belonged to P[13],I5 and G11,respectively.The virus was designated Rotavirus A pig/China/BJ/2015/G11P[13].
Rotavirus infections are a major cause of viral diarrheas in young animals and children.Isolation and identification of rotavirus make a contribution to epidemiological survey and molecular biology study.A strain of porcine rotavirus was isolated in MA104 cell cultures from the intestinal contents of piglets with diarrhea in Beijing.The virus was identified to be porcine rotavirus by immunochromatography strip test,Real-time RT-PCR,immunofluorescence test and sequencing analysis.According to the sequence analysis,the virus was classified as group A porcine rotavirus,the genotype of VP4,VP6 and VP7genes belonged to P[13],I5 and G11,respectively.The virus was designated Rotavirus A pig/China/BJ/2015/G11P[13].
引文
[1]Zimmerman J J.猪病学[M].第9版.北京:中国农业大学出版社,2008.
[2]Mebus C A,Underdahl N R,Rhodes M B,et al.Further studies on neonatal calf diarrhea virus[J].Proc Annu Meet U S Anim Health Assoc,1969,73:97-99.
[3]Bishop R F,Davidson G P,Holmes I H,et al.Virus particles in epithelial cells of duodenal mucosa from children with acute non-bacterial gastroenteritis[J].Lancet,1973,2(7841):1281-1283.
[4]Rodger S M,Craven J A,Williams I.Letter:Demonstration of reovirus-like particles in intestinal contents of piglets with diarrhoea[J].Aust Vet J,1975,51(11):536.
[5]He B,Yang F,Yang W,et al.Characterization of a novel G3P
[3]rotavirus isolated from a lesser horseshoe bat:A distant relative of feline/canine rotaviruses[J].J Virol,2013,87(22):12357-12366.
[6]Kindler E,Trojnar E,Heckel G,et al.Analysis of rotavirus species diversity and evolution including the newly determined full-length genome sequences of rotavirus F and G[J].Infect Genet Evol,2013,14:58-67.
[7]Desselberger U.Rotaviruses[J].Virus Research,2014,190:75-96.
[8]郑龙龙,朱玲云,陈关雄,等.断奶仔猪腹泻病因分析[J].中国畜牧兽医,2015,42(10):2772-2778.
[9]Martella V,Pratelli A,Elia G,et al.Isolation and genetic characterization of two G3P5A
[3]canine rotavirus strains in Italy[J].J Virol Methods,2001,96(1):43-49.
[10]Varghese V,Das S,Singh N B,et al.Molecular characterization of a human rotavirus reveals porcine characteristics in most of the genes including VP6and NSP4[J].Arch Virol,2004,149(1):155-172.
[11]Laird A R,Ibarra V,Ruiz-Palacios G,et al.Unexpected detection of animal VP7genes among common rotavirus strains isolated from children in Mexico[J].J ClinMicrobiol,2003,41(9):4400-4403.
[12]Esona M D,Armah G E,Geyer A,et al.Detection of an unusual human rotavirus strain with G5P[8]specificity in a Cameroonian child with diarrhea[J].J Clin Microbiol,2004,42(1):441-444.
[13]杨文宇,朱玲,周远成,等.猪轮状病毒四川株的分离鉴定及增殖规律[J].中国兽医学报,2014,34(2):192-198.
[14]陈晓春,吴华伟,张广川,等.猪轮状病毒的分离与鉴定[J].中国兽药杂志,2013,47(9):4-8.
[15]库旭钢,张坤,刘羽茜,等.猪A群轮状病毒的分离与鉴定[J].畜牧兽医学报,2012,43(2):275-281.
[16]时洪艳,陈建飞,王承宝,等.猪轮状病毒Rotavirus A pig/China/NMTL/2009/G9P
[23]株的分离与鉴定[J].中国预防兽医学报,2011,33(9):681-684.
[17]田小艳,孙华,邓雨修,等.猪轮状病毒分离与鉴定[J].畜牧与兽医,2009,41(10):85-87.
[18]王振玲,章丽娇,段跃强,等.猪轮状病毒的分离与电镜观察[J].畜牧与兽医,2014,46(1):77-80.
[19]张贺伟,王鑫,夏铭崎,等.猪A群轮状病毒LN-01-2013株的分离与鉴定[J].中国兽医科学,2014,5:470-475.
[20]夏乐乐,何彪,胡挺松,等.云南蝙蝠轮状病毒的分离与鉴定[J].病毒学报,2013,6:632-637.
[21]Matthijnssens J,Otto P H,Ciarlet M,et al.VP6-sequence-based cutoff values as a criterion for rotavirus species demarcation[J].Arch Virol,2012,157(6):1177-1182.
[22]Bohl E H,Theil K W,Saif L J.Isolation and serotyping of porcine rotaviruses and antigenic comparison with other rotaviruses[J].J Clin Microbiol,1984,19(2):105-111.
[23]Bautista D,Rodriguez L S,Franco M A,et al.Caco-2cells infected with rotavirus release extracellular vesicles that express markers of apoptotic bodies and exosomes[J].Cell Stress Chaperones,2015,20(4):697-708.
[24]范秀娟,孙燕,孙振鹏,等.生物反应器培养轮状病毒基因重配株Ls条件的优化[J].微生物学免疫学进展,2015,6:18-21.
[25]Burns J W,Greenberg H B,Shaw R D,et al.Functional and topographical analyses of epitopes on the hemagglutinin(VP4)of the simian rotavirus SA11[J].J Virol,1988,62(6):2164-2172.
[26]Papp H,Laszlo B,Jakab F,et al.Review of group A rotavirus strains reported in swine and cattle[J].Vet Microbiol,2013,165(3-4):190-199.
[2]Mebus C A,Underdahl N R,Rhodes M B,et al.Further studies on neonatal calf diarrhea virus[J].Proc Annu Meet U S Anim Health Assoc,1969,73:97-99.
[3]Bishop R F,Davidson G P,Holmes I H,et al.Virus particles in epithelial cells of duodenal mucosa from children with acute non-bacterial gastroenteritis[J].Lancet,1973,2(7841):1281-1283.
[4]Rodger S M,Craven J A,Williams I.Letter:Demonstration of reovirus-like particles in intestinal contents of piglets with diarrhoea[J].Aust Vet J,1975,51(11):536.
[5]He B,Yang F,Yang W,et al.Characterization of a novel G3P
[3]rotavirus isolated from a lesser horseshoe bat:A distant relative of feline/canine rotaviruses[J].J Virol,2013,87(22):12357-12366.
[6]Kindler E,Trojnar E,Heckel G,et al.Analysis of rotavirus species diversity and evolution including the newly determined full-length genome sequences of rotavirus F and G[J].Infect Genet Evol,2013,14:58-67.
[7]Desselberger U.Rotaviruses[J].Virus Research,2014,190:75-96.
[8]郑龙龙,朱玲云,陈关雄,等.断奶仔猪腹泻病因分析[J].中国畜牧兽医,2015,42(10):2772-2778.
[9]Martella V,Pratelli A,Elia G,et al.Isolation and genetic characterization of two G3P5A
[3]canine rotavirus strains in Italy[J].J Virol Methods,2001,96(1):43-49.
[10]Varghese V,Das S,Singh N B,et al.Molecular characterization of a human rotavirus reveals porcine characteristics in most of the genes including VP6and NSP4[J].Arch Virol,2004,149(1):155-172.
[11]Laird A R,Ibarra V,Ruiz-Palacios G,et al.Unexpected detection of animal VP7genes among common rotavirus strains isolated from children in Mexico[J].J ClinMicrobiol,2003,41(9):4400-4403.
[12]Esona M D,Armah G E,Geyer A,et al.Detection of an unusual human rotavirus strain with G5P[8]specificity in a Cameroonian child with diarrhea[J].J Clin Microbiol,2004,42(1):441-444.
[13]杨文宇,朱玲,周远成,等.猪轮状病毒四川株的分离鉴定及增殖规律[J].中国兽医学报,2014,34(2):192-198.
[14]陈晓春,吴华伟,张广川,等.猪轮状病毒的分离与鉴定[J].中国兽药杂志,2013,47(9):4-8.
[15]库旭钢,张坤,刘羽茜,等.猪A群轮状病毒的分离与鉴定[J].畜牧兽医学报,2012,43(2):275-281.
[16]时洪艳,陈建飞,王承宝,等.猪轮状病毒Rotavirus A pig/China/NMTL/2009/G9P
[23]株的分离与鉴定[J].中国预防兽医学报,2011,33(9):681-684.
[17]田小艳,孙华,邓雨修,等.猪轮状病毒分离与鉴定[J].畜牧与兽医,2009,41(10):85-87.
[18]王振玲,章丽娇,段跃强,等.猪轮状病毒的分离与电镜观察[J].畜牧与兽医,2014,46(1):77-80.
[19]张贺伟,王鑫,夏铭崎,等.猪A群轮状病毒LN-01-2013株的分离与鉴定[J].中国兽医科学,2014,5:470-475.
[20]夏乐乐,何彪,胡挺松,等.云南蝙蝠轮状病毒的分离与鉴定[J].病毒学报,2013,6:632-637.
[21]Matthijnssens J,Otto P H,Ciarlet M,et al.VP6-sequence-based cutoff values as a criterion for rotavirus species demarcation[J].Arch Virol,2012,157(6):1177-1182.
[22]Bohl E H,Theil K W,Saif L J.Isolation and serotyping of porcine rotaviruses and antigenic comparison with other rotaviruses[J].J Clin Microbiol,1984,19(2):105-111.
[23]Bautista D,Rodriguez L S,Franco M A,et al.Caco-2cells infected with rotavirus release extracellular vesicles that express markers of apoptotic bodies and exosomes[J].Cell Stress Chaperones,2015,20(4):697-708.
[24]范秀娟,孙燕,孙振鹏,等.生物反应器培养轮状病毒基因重配株Ls条件的优化[J].微生物学免疫学进展,2015,6:18-21.
[25]Burns J W,Greenberg H B,Shaw R D,et al.Functional and topographical analyses of epitopes on the hemagglutinin(VP4)of the simian rotavirus SA11[J].J Virol,1988,62(6):2164-2172.
[26]Papp H,Laszlo B,Jakab F,et al.Review of group A rotavirus strains reported in swine and cattle[J].Vet Microbiol,2013,165(3-4):190-199.