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A colloidal gold test strip assay for the detection of African swine fever virus based on two monoclonal antibodies against P30

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Abstract

African swine fever (ASF), caused by African swine fever virus (ASFV), was first reported in Kenya in 1921, but an effective vaccine or antiviral drug is still not available for ASFV control. Rapid and effective diagnostics are key steps in managing ASF. We generated two monoclonal antibodies (MAbs) against the ASFV phosphoprotein P30 and designated these as 3H7A7 and 6H9A10. Epitope mapping revealed that MAb 3H7A7 and 6H9A10 recognized aa 144-154 and aa 12-18 of P30, respectively. A signal-amplified sandwich colloidal gold test strip for rapid detection of ASFV was developed based using these MAbs. Sensitivity and specificity analysis showed that the detection limit of the strip was 2.16 ng of P30. The strip only reacted with ASFV and did not react with other common porcine viruses. In detection tests using 153 clinical field samples including sera, plasma, anticoagulant-treated blood, and tissue, the strip had 95.42% concordance with real-time PCR. The new MAbs specific for P30 and the rapid colloidal gold test strip helped to reveal novel B cell epitopes in P30 and provide an efficient diagnostic test for on-site clinical detection of ASF.

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Funding

This research was funded by the National Project for Prevention and Control of Transboundary Animal Diseases (Grant no. 2017YFD0501805), the National Key R&D Program for the 13th Five-Year Plan, and the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions.

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Authors and Affiliations

  1. College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, Jiangsu, China

    Xinyu Zhang, Xiaoyu Liu, Xiaoli Xia & Huaichang Sun

  2. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, Jiangsu, China

    Xinyu Zhang, Xiaoyu Liu, Xiaoli Xia & Huaichang Sun

  3. National Exotic Animal Disease Center, China Animal Health and Epidemiology Center, Qingdao, 266032, Shandong, China

    Xiaodong Wu, Weijie Ren & Yanli Zou

Authors
  1. Xinyu Zhang
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  2. Xiaoyu Liu
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  3. Xiaodong Wu
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  4. Weijie Ren
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  5. Yanli Zou
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  6. Xiaoli Xia
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  7. Huaichang Sun
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Contributions

XZ and XL performed most of the work; XW provided the field samples, the ASFV SY18 strain, and workspace in the biosafety level III laboratory; WR and YZ were responsible for testing field samples; XX helped in material preparation; and HS gave experimental instruction.

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Correspondence to Xinyu Zhang.

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All the authors declare that they have no competing interests regarding the publication of the data from this study.

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Handling Editor: Patricia Aguilar.

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Zhang, X., Liu, X., Wu, X. et al. A colloidal gold test strip assay for the detection of African swine fever virus based on two monoclonal antibodies against P30. Arch Virol 166, 871–879 (2021). https://doi.org/10.1007/s00705-020-04915-w

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  • DOI: https://doi.org/10.1007/s00705-020-04915-w

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