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Serodiagnosis, targeting nonstructural protein 4, of porcine reproductive and respiratory syndrome virus

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Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) causes great economic losses to the swine industry worldwide. Typically, an N protein-coated indirect enzyme-linked immunosorbent assay (N-coated iELISA) is used to detect PRRSV antibodies. Non-structural protein (NSP) 4 is essential to the PRRSV life cycle and contains B-cell epitopes. Yet, no specific antibody against NSP4 has been detected in clinical samples. In this study, we developed an NSP4-coated iELISA and compared its effectiveness with the N-coated iELISA. The NSP4-coated iELISA was developed with a cut-off value of 0.406 at an optical density of 450 nm by testing a panel of 70 PRRSV positive and 80 PRRSV negative pig serum samples, which generated a specificity and sensitivity of 100%. Agreement between the NSP4-coated and N-coated iELISAs was 92.2%. Interestingly, 50 serum samples, mostly from pigs vaccinated with the HP-PRRSV live strain, tested positive for PRRSV antibodies with the NSP4-coated iELISA, but were negative with the N-coated iELISA. These results were further confirmed by western blot analysis and another iELISA based on the N-terminus of NSP2 (NSP2-1-coated iELISA). The agreement between the results of western blot analysis with the NSP4-coated and NSP2-1-coated iELISA analyses were 92% and 96.1%, respectively, showing that the developed NSP4-coated iELISA is a useful tool to discriminate a false negative from a true negative response to the HP-PRRSV vaccine.

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Authors and Affiliations

  1. Department of Fundamental Veterinary Medicine, College of Animal Science and Veterinary Medicine, Shandong Agricultural University, 61 Daizong Street, Tai’an, 271018, Shandong, China

    Shengliang Cao, Xinna Cai, Min Tan, Fachao Sun, Yuchao Wang, Sidang Liu & Yihong Xiao

  2. Department of Preventive Veterinary Medicine, College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai’an, Shandong, China

    Yingli Shang

  3. Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Shandong Agricultural University, Tai’an, Shandong, China

    Shengliang Cao, Xinna Cai, Min Tan, Fachao Sun, Yuchao Wang, Yingli Shang, Sidang Liu & Yihong Xiao

Authors
  1. Shengliang Cao
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  2. Xinna Cai
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  3. Min Tan
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  4. Fachao Sun
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  5. Yuchao Wang
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  6. Yingli Shang
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  7. Sidang Liu
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  8. Yihong Xiao
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Corresponding author

Correspondence to Yihong Xiao.

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Funding

This study was funded by the National Natural Science Foundation of China (Grant no. 31772708), the Natural Science Foundation of Shandong Province (Grant no. ZR2014CM024) and the Funds of Shandong “Double Tops” Program.

Conflict of interest

The authors have no conflicts of interest to declare.

Ethical approval

All samples in this study were handled in accordance with the guidelines of the Animal Care and Use Committee of Shandong Agricultural University. This study did not involve human participants.

Additional information

Handling Editor: Zhenhai Chen.

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Cao, S., Cai, X., Tan, M. et al. Serodiagnosis, targeting nonstructural protein 4, of porcine reproductive and respiratory syndrome virus. Arch Virol 163, 411–418 (2018). https://doi.org/10.1007/s00705-017-3625-5

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  • DOI: https://doi.org/10.1007/s00705-017-3625-5

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