Vet World   Vol.18   August-2025  Article - 32 

Development of a cost-effective serodiagnosis for African swine fever using solubility-enhanced recombinant p54, p30, and p72

Background and Aim: The rapid spread of African swine fever (ASF) in Indonesia and other Asian countries has devastated domestic and wild pig populations. In the absence of a viable vaccine, ASF control depends on strict biosecurity measures and the prompt culling of infected animals. Accurate and timely detection is therefore essential to limit disease transmis­sion, highlighting the urgent need for reliable diagnostic tools. This study aimed to develop serological assays for ASF virus (ASFV) antibody detection using recombinant ASFV proteins.

Materials and Methods: Three key ASFV structural proteins–p30, p54, and p72–were heterologously expressed in Escherichia coli. Protein solubility, particularly for p54, was enhanced by targeted deletion of hydrophobic domains. Recombinant proteins were purified using nickel-nitrilotriacetic acid affinity chromatography and assessed for diagnostic performance through enzyme-linked immunosorbent assay (ELISA) and immunoblotting using 114 field serum samples.

Results: The solubility-optimized p54 antigen was successfully used to develop an indirect ELISA, while the insoluble p30 retained sufficient antigenicity for immunoblot-based detection. The p54-based ELISA showed high diagnostic performance, achieving an area under the curve of 0.936, with 91% sensitivity and 85% specificity. Agreement with a commercial ELISA kit was substantial (Cohen’s kappa = 0.635). Immunoblotting confirmed that all recombinant proteins maintained strong antigenicity and diagnostic specificity.

Conclusion: Recombinant ASFV proteins p54 and p30 demonstrated strong potential for serological diagnostics when expressed in E. coli. Notably, this is the first study to report a successful domain truncation strategy for enhancing p54 solu­bility in E. coli, enabling the development of affordable, locally produced ELISA kits. The p30-based immunoblot assay serves as a confirmatory tool to strengthen ASF detection and outbreak response in resource-limited settings.

Keywords: African swine fever, enzyme-linked immunosorbent assay, recombinant p30, recombinant p54, recombinant p72, serodiagnosis.

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How to cite this article: Tarigan S, Sumarningsih S, Ratnawati A, Saepulloh M, Wasito W, Sendow I, Nuradji H, and Dharmayanti NLPI (2025) Development of a cost-effective serodiagnosis for African swine fever using solubility-enhanced recombinant p54, p30, and p72, Veterinary World, 18(8): 2511-2519.

Received: 24-04-2025   Accepted: 28-07-2025   Published online: 30-08-2025

Corresponding author: Simson Tarigan    E-mail: sitariganta@gmail.com

DOI: 10.14202/vetworld.2025.2511-2519

Copyright: Tarigan, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.