ISSN: 0973-7510

E-ISSN: 2581-690X

Research Article | Open Access
Manjunatha Jogaiah1,2, Sharanagouda S. Patil1 , Pavithra Narendran2, Kuralayanapalya Puttahonnappa Suresh1 and Chandranaik Basavegowdanadoddi Marinaik3
1ICAR-National Institute of Veterinary Epidemiology and Disease Informatics, Yelahanka, Bengaluru, Karnataka, India.
2The University of Trans-Disciplinary Health Sciences and Technology (TDU), Jarakabande Kaval, Yelahanka, Bengaluru, India.
3Institute of Animal Health and Veterinary Biologicals, Karnataka Veterinary, Animal and Fisheries Sciences University (KVAFSU), Hebbal, Bengaluru, Karnataka, India.
Article Number: 11426 | © The Author(s). 2026
J Pure Appl Microbiol. 2026;20(3):2112-2124. https://doi.org/10.22207/JPAM.20.3.02
Received: 10 February 2026 | Accepted: 21 May 2026 | Published online: 01 July 2026
Issue online: September 2026
Abstract

Classical Swine Fever (CSF) remains a highly transmissible viral threat to the Suidae family with considerable economic consequences worldwide. The current study developed and comprehensively validated an indirect ELISA that uses the recombinant E2 protein (rE2) from CSFV. The amplified 546 bp segment from the N-terminal of the CSFV E2 gene, cloned into the pET-28a vector, and introduced into E. coli TOP10 and subsequently into BL21(DE3) cells. A ~25 kDa His-tagged rE2 protein was successfully expressed, followed by solubility analysis and purified using affinity chromatography. The recombinant protein was verified by Western blot analysis employing anti-His antibodies and CSFV positive pig sera. Using 500 pig serum samples, the developed iELISA diagnostic performance was evaluated and compared with both an indigenously developed Erns ELISA kit and Priocheck CSFV Antibody ELISA kit. ROC analysis of developed rE2 iELISA demonstrated robust diagnostic precision, yielding an area under the curve (AUC) value of 0.935 (92% sensitivity, 90% specificity; cut-off 0.5695) against the Priocheck CSFV Antibody ELISA kit and with indigenously developed Erns ELISA assay (AUC 0.995; 97.98% sensitivity; 96.97% specificity at the cut-off of 0.404). A statistically significant variation in diagnostic accuracy of two assays was evident in comparative ROC curve analysis (AUC difference = 0.216; SE = 0.0482; 95% CI: 0.122-0.311; z = 4.484; P < 0.0001) proving the developed assay provides a sensitive, specific, and robust serological tool suitable for CSF serosurveillance.

Keywords

Classical Swine Fever Virus (CSFV), Recombinant E2 Protein (rE2 Protein), Indirect Enzyme Linked Immuno Sorbent Assay (iELISA), Serological Diagnosis, Diagnostic sensitivity and specificity, ROC Analysis, Immunomonitoring

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© The Author(s) 2026. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, 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.