PRRSV河北分离株ORF5与NSP2基因遗传变异分析
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摘要
从河北省新乐市某猪场采集疑似感染猪繁殖与呼吸综合征病毒(PRRSV)的病料,通过Marc-145细胞培养,RT-PCR检测等方法鉴定,确认分离到1株PRRSV,暂命名为HB-XL株。进一步对其ORF5与NSP2基因克隆测序,经同源性比较及系统进化树等分析,表明该毒株ORF5与NSP2基因与国内流行的缺失变异株遗传关系较近,与国内外经典美洲型毒株及基因重组毒株QYYZ遗传关系较远且与欧洲型LV毒株处于不同分支;ORF5基因编码的200个氨基酸的第13、151位均为具有强毒特性的精氨酸(R),第137位为具有野毒特性的丝氨酸(S);与经典美洲株相比,该毒株NSP2基因分别发生3个碱基和87个碱基的不连续缺失。说明HB-XL株属于美洲型PRRSV并存在不断变异的趋势。
A porcine productive and respiratory syndrome virus(PRRSV)field strain,tentatively named HB-XL,was isolated from suspicious PRRSV-positive materials of sick pigs in Xinle areas of Hebei province,according to the Marc-145 cell culture and RT-PCR detection method.Further analysis by cloning ORF5 and NSP2sequencing,homology comparison and phylogenetic evolutionary tree showed that the ORF5 and NSP2of HB-XL share the similar mutant genetic relationship to the majority of the domestic epidemic PRRSV strains,and the relatively distant relationship to the American classical strains and recombinant strain QYYZ,and the Europe strain LV is an independent branch;The ORF5 gene consisted of 200 amino acids and the sites of 13 and 151were argrinine(R)which were related to virulent strains,and the site of 137 was serine(S)indicating that the isolated strain was wild strain;the Nsp2 gene had a discontinuous deletions of 3bases and 87 bases compared with the American classical strains.The results showed that PRRSV HB-XL strain belongs to the American type PRRSV,and with a trend of continuous variation.
A porcine productive and respiratory syndrome virus(PRRSV)field strain,tentatively named HB-XL,was isolated from suspicious PRRSV-positive materials of sick pigs in Xinle areas of Hebei province,according to the Marc-145 cell culture and RT-PCR detection method.Further analysis by cloning ORF5 and NSP2sequencing,homology comparison and phylogenetic evolutionary tree showed that the ORF5 and NSP2of HB-XL share the similar mutant genetic relationship to the majority of the domestic epidemic PRRSV strains,and the relatively distant relationship to the American classical strains and recombinant strain QYYZ,and the Europe strain LV is an independent branch;The ORF5 gene consisted of 200 amino acids and the sites of 13 and 151were argrinine(R)which were related to virulent strains,and the site of 137 was serine(S)indicating that the isolated strain was wild strain;the Nsp2 gene had a discontinuous deletions of 3bases and 87 bases compared with the American classical strains.The results showed that PRRSV HB-XL strain belongs to the American type PRRSV,and with a trend of continuous variation.
引文
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[15]刘建奎,魏春华,杨小燕,等.福建地区PRRSV分离株Nsp2基因新变异序列鉴定[J].中国预防兽医学报,2013,35(4):271-275.
[16]Ni Y Y,Huang Y W,Cao D,et al.Establishment of a DNA-launched infectious clone for a highly pneumovirulent strain of type 2porcine reproductive and respiratory syndrome virus:identification and in vitro and in vivo characterization of a Large spontaneous deletion in the nsp2region[J].Virus Res,2011,160(12):264-273.
[2]郭宝清,陈章水,刘文兴,等.从疑似PRRS流产胎儿分离PRRSV的研究[J].中国畜禽传染病,1996(2):1-4.
[3]童光志,周艳君,郝晓芳,等.高致病性猪生殖与呼吸综合征病毒的分离鉴定及其分子流行病学分析[J].中国预防兽医学报,2007,27(5):333-336.
[4]Mateu E,Díaz I,Darwich L,et al.Evolution of ORF5of Spanish porcine reproductive and respiratory syndrome virus strains from 1991to 2005[J].Virus Res,2006,115(2):198-206.
[5]Kim W I,Kim J J,Cha S H,et al.Different biological characteristics of wild-type porcine reproductive and respiratory syndrome viruses and vaccine viruses and identification of the corresponding genetic determinants[J].Clin Microbiol,2008,46(5):1758-1768.
[6]童光志,周艳君,都晓芳,等.高致病性猪繁殖与呼吸综合征病毒的分离鉴定及其分子流行病学分析[J].中国预防兽医学报,2007,29(5):323-327.
[7]Key K F,Dicristina J,Gillespie J,et al.Direct inoculation of RNA transcripts from an infectious cDNA clone of porcine reproductive and respiratory syndrome virus(PRRSV)into the lymph nodes and tonsils of pigs initiates PRRSV infection in vivo[J].Arch Virol,2007,152(7):1383-1387.
[8]周铁忠,王占林,刘丽颖,等.PRRSV LN/0901株的分离鉴定及其NSP2基因遗传变异分析[J].中国兽医学报,2012,32(5):1758-1762.
[9]Zongxi C,Quanhui P,Zhemin L,et al.Isolation and sequence analysis of NSP2,ORF3 and ORF5 of PRRSV XH-GD strain from Guangdong province[J].Agricul Biotechnol,2014,3(1):4-8.
[10]姚敬明,刘文俊,罗甜甜,等.PRRSV山西株ORF5和NSP2基因序列分析[J].中国兽医学报,2014,34(1):1-5.
[11]Jiankui L,Chunhua W,Xiaoyan Y,et al.Genetic diversity and evolutionary characterization of Chinese porcine reproductive and respiratory syndrome viruses based on NSP2and ORF5[J].Arch Virol,2013,158(8):1811-1816.
[12]陈家锃,白云,常丹,等.一株不能被高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)疫苗抗血清中和的HP-PRRSV的分离鉴定[J].中国预防兽医学报,2014,36(3):174-177.
[13]Yuhang C,Hongsheng Y,Mingjun Z H,et al.Analysis of molecular variation in porcine reproductive and respiratory syndrome virus in China between 2007and 2012[J].Virol Sinica,2014,29(3):183-188.
[14]Bin L,Liurong F,Xueliang G,et al.Epidemiology and evolutionary characteristics of the porcine reproductive and respiratory syndrome virus in China between2006and 2010[J].Clin Microl,2011,49(9):3175-3183.
[15]刘建奎,魏春华,杨小燕,等.福建地区PRRSV分离株Nsp2基因新变异序列鉴定[J].中国预防兽医学报,2013,35(4):271-275.
[16]Ni Y Y,Huang Y W,Cao D,et al.Establishment of a DNA-launched infectious clone for a highly pneumovirulent strain of type 2porcine reproductive and respiratory syndrome virus:identification and in vitro and in vivo characterization of a Large spontaneous deletion in the nsp2region[J].Virus Res,2011,160(12):264-273.