Isolation,Identification and Genetic Variation of Porcine Reproductive and Respiratory Syndrome Virus Variant

doi:10.7668/hbnxb.2014.01.042

Abstract

Abstract: A porcine reproductive and respiratory syndrome virus(PRRSV) strain was isolated from the serum of a PRRSV infected pig in Jiangsu Province. The biological characteristics of this isolate was identified and the genome of Nsp2 and ORF5 were determined and analyzed. The serum which were detected PRRSV positive and PPV, PRV negative was inoculated and passaged on Marc-145 cells. This isolate was identified by RT-PCR and CPE,and confirmed by immunofluorescence assay(IFA). The TCID ₅₀ of this isolate was measured and the genome of Nsp2 and ORF5 were sequenced and its homology and evolutionary relationship were analyzed by DNAStar software. The results indicated that the PRRSV strain was successfully isolated and it could induce cytopathogenic effect(CPE) on Marc-145 cells. The specific fluorescence could be observed and the TCID ₅₀ of this strain was 10^{5. 5}. Comparison analysis of the Nsp2 genome of the isolate 2010JS02 and other strains revealed that the isolate has a discontinuous deletion of 30 amino acids and another deletion at the position of 495- 516 amino acid's site. According to the se-quences analysis of GP5,the isolate 2010JS02 showed identity of 57. 1%- 99. 5% among other strains,a high identity of 99. 5% with NJ-1106. The phylogenetic tree was combined and showed that 2010JS02 belonged to NA and classified into HP-PRRSV like WHU4,NJ-1106 and other HP-PRRSV strains. We successfully isolated a HPPRRSV variant and named it as 2010JS02. These results provided foundations for further studying the genetic variation and the prevention of PRRSV.

Key words: Porcine reproductive and respiratory syndrome virus(PRRSV), Isolation and identification, Genet-is vanatron

CLC Number:

S432.4

WANG Xiao-min, HE Kong-wang, ZHOU Zhong-tao, MAO Ai-hua, YU Zheng-yu, WANG Wei, NI Yan-xiu, WEN Li-bin, ZHANG Xue-han, GUO Rong-li, LI Bin, YU Yang. Isolation,Identification and Genetic Variation of Porcine Reproductive and Respiratory Syndrome Virus Variant[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(1): 232-238. doi: 10.7668/hbnxb.2014.01.042.

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References

1 Zimmerman J J, Yoon K J,Wills RW,et al. General over-view of PRRSV:a perspecaive from the United States[J]. Microbio1,1997,55(1-4):187-196
2 Rossow K D. Porcine reproducaive and respiratory syn-drome[J]. Vet Pathol,1998, 35:1-20
3 Kelfaver K K. Reproducaive failure of unknown etiology[J]. Am Assoc Swine Praca News，1989，1(2):1-9
4 Nelsen C J, Murtaugh M P, Faaberg K S. Porcine repro-ducaive and respiratory syndrome virus comparison:diver-gent evolution on two continents[J]J Virol, 1999，73(1):270-80.
5 Meulenberg J J,de Meijer E J,Moormann R J. Subgenom-ic RNAs of Lelystad virus。ontain a conserved leader-body juncaion sequence[J].J Gen Viro1,1993,74(8):1697-1701
6 Stadejek T, Oleksiewicz M B，Potapc:huk D, et al. Porcine reproducaive and respiratory syndrome virus strains of ex-ceptional diversity in eastern Europe support the definition of new genetic: subtypes [J]. J Gen Virol,2006, 87 (Pt7): 1835-1841
7 Collins J E，Benfield D A, Christianson WT, et al. Isola-tion of swine infertility and respiratory syndrome virus (i-solute ATCC VR-2332) in North America and experimen-tal reproducaion of the disease in gnotobiotic pigs[J].J Vet Diagn,1992,4(2):117-126
8 郭宝清，陈章水，刘文兴，等从疑似PRRSV感染的流产胎儿分离猪生殖和呼吸综合征病毒(PRRSV)的研究[J]中国畜禽传染病，1996,87(2);1 -5
9 宋凌百，文英会，崔保安，等猪繁殖与呼吸综合征在我国的流行动态[J]上海畜牧兽医通讯，2005 (2); 43-45
10 Tian K G, Yu X L, Zhao T Z，et al. Emergence of fatal PRRSV variants:unparalleled outbreaks of atypical PRRS in China and molecular dissecaion of the unique hallmark[J]. P1oS ONE,2007,2(6);526-539
11 Tong G Z, Zhou Y J, Hao X F, et al. Highly pathogenic porcine reproducaive and respiratory syndrome, China [J].Emerging Infecioust Diseases,2007，13(9):1434-1436
12 Zhou L, Yang H C. Porcine reproducaive and respiratory syndrome in China[J]. Virus Res,2010，154(1-2):31-37
13 安同庆，田志军，李冉，等我国高致病性猪蓝耳病病毒的演化分析[J]中国兽医杂志，2011,47(4);3-6
14 王小敏，何孔旺，张文文，等2008-2010年中国华东地区PRRSV ORFS基因遗传变异分析[J]中国农业科学，2011,44(18):3886-3894
15 Den Boon J A,Faaherg K S,Meulenherg J J, et al. Pro-cessing and evolution of the N-terminal region of the ar-terivirus replicase ORFla protein:identification of twoPapainlike cysteine proteases[J]. Virol, 1995，69(7):4500-4505
16 Li Y,Wang X,Bo K,et al. Emergence of a highly patho-genic porcine reproducaive and respiratory syndrome vi-rus in the Mid-Eastern region of China[J]. Vet J,2007，174(3);577-584
17 Zhou L,Zhang J,Zeng J,et al. The 30-amino-acid deletionin the Nsp2 of high1y pathogenic: porcine reproducaive and respiratory syndrome virus emerging in China is not relat-ed to its virulence[J]I Virol, 2009，83(10):5156-5157
18 Zhou Y J, Yu H, Tian Z J, et al. Genetic: diversity of theORFS gene of porcine reproducaive and respiratory syn-drome virus isolates in China from 2006 to 2008[J]Virus Res,2009，144(1-2):136-144
19 Yin G, Gan L, Shu X, et al. Genetic: diversity of theORFS gene of porcine reproducaive and respiratory syn-drome virus isolates in southwest China from 2007 to2009[J]. Plos One,2012,7(3):e33756
20 Wei X,Decker J M，Wang S,et al. Antibody neutraliza-tion and escape by HIV-1[J]Nature, 2003，422(6929);307-312
21 Han J,Wang Y,Faaberg K S. Complete genome analysisof RFLP 184 isolates of porcine reproducaive and respira-tort' syndrome virus[J]. Virus Res,2006，122(1-2):175-182.

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