Establishment of a Dual-Antigen Indirect ELISA Based on p30 and pB602L to Detect Antibodies against African Swine Fever Virus

Lei Zhou; Jinxing Song; Mengxiang Wang; Zhuoya Sun; Junru Sun; Panpan Tian; Guoqing Zhuang; Angke Zhang; Yanan Wu; Gaiping Zhang

doi:10.3390/v15091845

Establishment of a Dual-Antigen Indirect ELISA Based on p30 and pB602L to Detect Antibodies against African Swine Fever Virus

Viruses. 2023 Aug 30;15(9):1845. doi: 10.3390/v15091845.

Authors

Lei Zhou^{1

2}, Jinxing Song^{1

2}, Mengxiang Wang^{1

2}, Zhuoya Sun^{1

2}, Junru Sun^{1

2}, Panpan Tian^{1

2}, Guoqing Zhuang^{1

2}, Angke Zhang^{1

2}, Yanan Wu^{1

2}, Gaiping Zhang^{1

2

3}

Affiliations

¹ International Joint Research Center of National Animal Immunology, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China.
² Ministry of Education Key Laboratory for Animal Pathogens and Biosafety, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China.
³ Longhu Laboratory of Advanced Immunology, Zhengzhou 450046, China.

Abstract

African swine fever (ASF) is an acute, virulent, and highly fatal infectious disease caused by the African swine fever virus (ASFV). There is no effective vaccine or diagnostic method to prevent and control this disease currently, which highlights the significance of ASF early detection. In this study, we chose an early antigen and a late-expressed antigen to co-detect the target antibody, which not only helps in early detection but also improves accuracy and sensitivity. CP204L and B602L were successfully expressed as soluble proteins in an Escherichia coli vector system. By optimizing various conditions, a dual-antigen indirect ELISA for ASFV antibodies was established. The assay was non-cross-reactive with antibodies against the porcine reproductive and respiratory syndrome virus, classical swine fever virus, porcine circovirus type 2, and pseudorabies virus. The maximum serum dilution for detection of ASFV-positive sera was 1:1600. The intra-batch reproducibility coefficient of variation was <5% and the inter-batch reproducibility coefficient of variation was <10%. Compared with commercial kits, the dual-antigen indirect ELISA had good detection performance. In conclusion, we established a detection method with low cost, streamlined production process, and fewer instruments. It provides a new method for the serological diagnosis of ASF.

Keywords: African swine fever virus; antibody detection; dual-antigen indirect ELISA; serological diagnosis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

African Swine Fever Virus*
African Swine Fever* / diagnosis
Animals
Antibodies
Enzyme-Linked Immunosorbent Assay
Escherichia coli
Reproducibility of Results
Swine

Substances

Antibodies

Grants and funding

This work was supported by the National Natural Science Foundation of China (NSFC) (32002292) and the Major Science and Technology Project of Henan Province (221100110600).