A Robust Quadruple Protein-Based Indirect ELISA for Detection of Antibodies to African Swine Fever Virus in Pigs

Author:

Jung Min-Chul12,Le Van Phan3ORCID,Yoon Sun-Woo4,Le Thi Ngoc12,Trinh Thi Bich Ngoc3,Kim Hye Kwon5,Kang Jung-Ah1,Lim Jong-Woo6,Yeom Minjoo6,Na Woonsung7ORCID,Nah Jin-Ju8,Choi Ji-Da8,Kang Hae-Eun8ORCID,Song Daesub6,Jeong Dae Gwin12ORCID

Affiliation:

1. Bionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Republic of Korea

2. Department of Proteome Structural Biology, KRIBB School of Bioscience, University of Science and Technology, Daejeon 34141, Republic of Korea

3. Department of Microbiology and Infectious Diseases, Faculty of Veterinary Medicine, Vietnam National University of Agriculture, Hanoi 100000, Vietnam

4. Department of Biological Science and Biotechnology, Andong National University, Andong 36729, Republic of Korea

5. Department of Microbiology, College of Natural Sciences, Chungbuk National University, Cheongju 28644, Republic of Korea

6. College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 08826, Republic of Korea

7. College of Veterinary Medicine, Chonnam National University, Gwangju 61186, Republic of Korea

8. Foreign Animal Disease Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea

Abstract

African swine fever (ASF) emerged in domestic pigs and wild boars in China in 2018 and rapidly spread to neighboring Asian countries. Currently, no effective vaccine or diagnostic tests are available to prevent its spread. We developed a robust quadruple recombinant-protein-based indirect enzyme-linked immunosorbent assay (QrP-iELISA) using four antigenic proteins (CD2v, CAP80, p54, and p22) to detect ASF virus (ASFV) antibodies and compared it with a commercial kit (IDvet) using ASFV-positive and -negative serum samples. The maximum positive/negative value was 24.033 at a single antigen concentration of 0.25 μg/mL and quadruple ASFV antigen combination of 1 μg/mL at a 1:100 serum dilution. Among 70 ASFV-positive samples, 65, 67, 65, 70, 70, and 14 were positive above the cut-offs of 0.121, 0.121, 0.183, 0.065, 0.201, and 0.122, for CD2v, CAP80, p54, p22-iELISA, QrP-iELISA, and IDvet, respectively, with sensitivities of 92.9%, 95.7%, 92.9%, 100%, 100%, and 20%, respectively, all with 100% specificity. The antibody responses in QrP-iELISA and IDvet were similar in pigs infected with ASFV I. QrP-iELISA was more sensitive than IDvet for early antibody detection in pigs infected with ASFV II. These data provide a foundation for developing advanced ASF antibody detection kits critical for ASF surveillance and control.

Funder

KRIBB Initiative Program

Korean government

Vietnamese government

Publisher

MDPI AG

Subject

Virology,Microbiology (medical),Microbiology

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