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Application of an AlphaLISA method for rapid sensitive detection of African swine fever virus in porcine serum

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Abstract

Infection with African swine fever virus (ASFV) causes an acute and highly lethal hemorrhagic disease that has been responsible for huge economic losses in China. To exactly detect the antigen of ASFV, we established a rapid, no-wash, one-step sandwich-type immunoassay based on the amplified luminescent proximity homogenous assay-linked immunosorbent assay (AlphaLISA) using two monoclonal antibodies (mAbs) M-5 and M-6 against ASFV p72. ASFV p72 in samples was captured by biotinylated mAb M-5 connected to the donor bead surface via streptavidin and “sandwiched” by mAb M-6 which was coated onto the acceptor bead. Efficacy and sensitivity trials revealed that the AlphaLISA could detect ≥0.78 ng/ml of purified p72 and with a linear range of 0.78–100 ng/ml. The AlphaLISA was specific for ASFV and did not cross-react with other common pathogenic porcine viruses. Compared with RealPCR ASFV DNA test and ASFV antigen detection kit, the sensitivity of the AlphaLISA evaluated in 60 porcine serum samples was 93% and 100%, respectively. The specificity was 100% and 91.7%, respectively. This study presents a good laboratory diagnostic tool for sensitive and efficient detection of ASFV in porcine serum.

Key points

MAbs M-5 and M-6 recognized various epitopes of ASFV p72.

The established ASFV p72 AlphaLISA showed well specificity, high sensitivity, and satisfied correlation coefficient.

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All data is presented in the manuscript, and the authors promise the availability of supporting data.

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Acknowledgements

We are grateful to Chunyan Feng (Institute of Animal Inspection and Quarantine, Chinese Academy of Inspection and Quarantine, Beijing, China) for the technical data analysis.

Funding

This work was supported by the National Science and Technology Support Programs of China (grant 2016YFD0501102), Key R&D Program of Shaanxi Province (2020NY-027), Ankang Leading Industry Major Research Project (AK2019ZD-10), and the fundamental research funds for CAIQ (grant 2020JK021).

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Contributions

SQ Wu conducted and designed the experiment. DJ Chen and D Wang performed research and wrote the paper, CX Wang and F Wei provided reagents and analyzed data, and HY Zhao provided the inactivated porcine serum and assisted in the ASFV antigen detection assay. XM Lin conducted the experiment.

Corresponding author

Correspondence to Shaoqiang Wu.

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Chen, D., Wang, D., Wang, C. et al. Application of an AlphaLISA method for rapid sensitive detection of African swine fever virus in porcine serum. Appl Microbiol Biotechnol 105, 4751–4759 (2021). https://doi.org/10.1007/s00253-021-11339-2

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  • DOI: https://doi.org/10.1007/s00253-021-11339-2

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