基于重组NS1蛋白猪细小病毒野毒抗体间接ELISA诊断方法的建立与应用

Abstract

Abstract: Using recombinant NS1 protein as coating antigen, an indirect ELISA was successfully developed to detect wild virus antibody to PPV. The results of detecting the positive sera of other five swine diseases(CSFV, PRRSV, PRV, PCV2, PEDV) using the method were negative. Coefficient of variability percent (C.V%) of intro-batch and inter-batch duplicative tests was less than 5% and 10%, respectively. Lowest antibody titer of positive control serum was 1:12800. Comparing with the HI assay, the concordance rate was 100%. Therefore, the NS1-ELISA had good specificity, sensitivity and repeatability, and could be used as a simple and rapid serology detection method for monitoring the anti-PPV wild virus antibody and epidemiologic survey of PPV.

Key words: porcine parvovirus; NS1 protein; indirect ELISA; diagnosis

CLC Number:

Q786

TANG Zhong-he. Establishment and Application of an Indirect ELISA for Detecting Wild Virus Antibodies to Porcine Parvovirus using NS1 Recombinant Protein[J]. China Animal Husbandry & Veterinary Medicine, 2013, 40(1): 46-49.

References

1 苏乾莲, 李斌, 赵武, 等.猪细小病毒自然弱毒N株非结构蛋白NS1基因原核表达[J].广西农业科学,2010,41(7):710~713.
2 陈光达,许信刚,童德文. PCV2、PPV和PRV多重PCR检测方法的建立及初步应用[J].西北农林科技大学学报(自然科学版),2011,39(11):1~6.
3 赵丹,贾红,侯绍华,等.犬细小病毒VP2抗体竞争ELISA检测方法的初步建立[J].中国畜牧兽医,2012,39(4):11~16.
4 夏应菊,訾占超,蔡林,等.猪细小病毒和猪圆环病毒2型混合感染状况的调查[J].畜牧兽医学报,2012,43(3):424~430.
5 殷华平,郭万柱,徐志文,等.猪细小病毒(PPV)SC1株非结构蛋白NS1基因的克隆和序列分析[J].中国预防兽医学报, 2005,27(6):504~508.
6 顾帅, 陈陆, 常洪涛, 等. SYBR GreenⅠ实时PCR对猪细小病毒体外复制动态分析[J].中国兽医学报,2012,32(3):350~354.
7 Cartwright S F, Huck R A. Virus isolation is associated with herd infertility, abortion and stillbirth in pigs[J].Vet Rec, 1967,81:196~197.
8 Cui J, Wang X, Ren Y, et al. Genome sequence of Chinese porcine parvovirus strain PPV2010[J]. J Virol, 2012,86(4):2379.
9 Cui S J, Martí C, Joaquim S. Phylogeny and evolution of the NS1 and VP1/VP2 gene sequences from porcine parvovirus[J]. Virus Res, 2009,140(1~2):209~215.
10 Qing L, Lv J, Li H, et al. The recombinant nonstructural polyprotein NS1 of porcine parvovirus (PPV) as diagnostic antigen in ELISA to differentiate infected from vaccinated pigs[J]. Vet Res Commun, 2006,30(2):175~190.
11 Sharma R, Saikumar G. Porcine parvovirus and porcine circovirus 2 associated reproductive failure and neonatal mortality in crossbred Indian pigs[J].Trop Anim Health Prod, 2010,42(3):515~522.
12 Song C, Zhu C, Zhang C, et al. Detection of porcine parvovirus using a TaqMan-based real-time PCR with primers and probe designed for the NS1 gene[J]. Virol J, 2010,7(1):353.
13 Tummaruk P, Tantilertcharoen R. Seroprevalence of porcine reproductive and respiratory syndrome, Aujeszky’s disease, and porcine parvovirus in replacement gilts in Thailand[J]. Trop Anim Health Prod, 2012,44(5):983~989.

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Establishment and Application of an Indirect ELISA for Detecting Wild Virus Antibodies to Porcine Parvovirus using NS1 Recombinant Protein

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