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The Recombinant Nonstructural Polyprotein NS1 of Porcine Parvovirus (PPV) as Diagnostic Antigen in ELISA to Differentiate Infected from Vaccinated Pigs

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Abstract

To differentiate pigs infected with porcine parvovirus (PPV) from those vaccinated with inactivated whole-virus vaccine, an enzyme-linked immunosorbent assay (ELISA) based on detection of a nonstructural polyprotein 1 (NS1) was developed. A threshold of 0.23 optical density units was established and the assayhad high specificity (100), sensitivity (88), accuracy (90) and positive predictive value (100) using haemagglutination inhibition as the standard method. A reproducibility test revealed that the coefficients of variation of sera within-plates and between-run were less than 10%. The assayshowed no cross-reactivitywith antibodies to porcine reproductive respiratorysyndrome virus, pseudorabies virus, foot and mouth disease virus, Actinobacillus pleuropneumoniae, Toxoplasma or Chlamydia. Sera obtained from pigs infected with PPV reacted with recombinant NS1 protein in the ELISA. Sera from pigs vaccinated with inactivated whole virus did not recognize this protein in the ELISA. In contrast, antibodies against PPV whole virus were present in both PPV-infected and vaccinated animals. Serum conversion against NS1 was first detected 10 days after infection and NS1-specific antibodies were detectable up to half a year post infection. In conclusion, the PPV-NS1 ELISA can differentiate PPV-infected versus inactivated PPV-vaccinated pigs and could be applied in disease diagnosis and surveillance.

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Abbreviations

ADV:

Aleutian mink disease parvovirus

APP:

Actinobacillus pleuropneumonia

CV:

coefficient of variation

ELISA:

enzyme-linked immunosorbent assay

FA:

fluorescent antibody

FMDV:

foot and mouth disease virus

HA:

haemagglutination

HI:

haemagglutination inhibition

HRP:

horseradish peroxidase

IgG:

immunoglobulin G

IPTG:

isopropyl-b-d -thiogalactopyranoside

MVM:

minute virus of mice

NS:

nonstructural

NT:

neutralization test

OD:

optical density

ORF:

open reading frame

PCR:

polymerase chain reaction

PCV2:

porcine circovirus 2

PMWS:

postweaning multisystemic wasting disease

PPV:

porcine parvovirus

PRRSV:

porcine reproductive and respiratory syndrome virus

PRV:

pseudorabies virus

SDS-PAGE:

sodium dodecyl sulphate– polyacrylamide gel electrophoresis

TCID50:

tissue culture infective dose 50%

TMB:

3,3,5,5-tetramethylbenzidine

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Authors and Affiliations

  1. Laboratory of Animal Virology, College of Veterinary Medicine, and The National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, P. R. China

    L. Qing, J. Lv, H. Li, Y. Tan, H. Hao, Z. Chen, J. Zhao & H. Chen

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  1. L. Qing
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  2. J. Lv
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  3. H. Li
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  4. Y. Tan
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  5. H. Hao
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  6. Z. Chen
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  7. J. Zhao
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  8. H. Chen
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Correspondence to H. Chen.

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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. Vet Res Commun 30, 175–190 (2006). https://doi.org/10.1007/s11259-006-3212-9

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  • DOI: https://doi.org/10.1007/s11259-006-3212-9

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