Cloning and Expression of a Truncated Form of the p72 Protein of the African Swine Fever Virus (ASFV) for Application in an Efficient Indirect ELISA System

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This research was funded by PROCINORTE Project 2023 (INIFAP-CENID-SAI SIGI 154626612) and the Spanish Ministry of Science and Innovation (grant reference PID2022-136312OB-I00 to F.R.).

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Pina-Pedrero, SoniaAuthorGanges, LlilianneAuthorRodriguez, FernandoAuthor

June 30, 2025

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Article

Publicated to:Pathogens. 14 (6): 542- - 2025-05-29 14(6), DOI: 10.3390/pathogens14060542

Authors: Cuevas-Romero, Julieta Sandra; Zavala-Ocampo, Perla Lucero; Pina-Pedrero, Sonia; Ganges, Llilianne; Muñoz-Aguilera, Adriana; Garcia-Cambron, Jose Bryan; Rodriguez, Fernando; Ambagala, Aruna; Cerriteño-Sanchez, Jose Luis

Affiliations

Campus Univ Autonoma Barcelona UAB, Ctr Recerca Sanitat Anim CReSA, IRTA, Anim Hlth, Bellaterra 08193, Spain - Author

Campus Univ Autonoma Barcelona UAB, Ctr Recerca Sanitat Anim CReSA, Unitat Mixta Invest IRTA UAB Sanitat Anim, Bellaterra 08193, Spain - Author

Inst Colombiano Agropecuario ICA, Bogota 110911, Colombia - Author

Inst Nacl Invest Forestales Agr & Pecuarias, Ctr Nacl Invest Disciplinaria Salud Anim Inocu, Mexico City 05110, Mexico - Author

Natl Ctr Foreign Anim Dis, Winnipeg, MB R3E 3M4, Canada - Author

Univ Autonoma Metrolitana Iztapalapa, Posgrad Biol Expt, Mexico City 09340, Mexico - Author

Univ Nacl Autonoma Mexico, Fac Med Vet, Posgrad Ciencias Prod & Salud Anim, Ave Univ 3004,Copilco Univ, Mexico City 04510, Mexico - Author

Abstract

African swine fever (ASF) is a disease that affects both domestic and wild swine. It was recently reported in the Dominican Republic and Haiti (2021), representing a substantial risk to America. The goal of this study was to produce a truncated form of the ASF-p72 recombinant protein based on the ASF strain genotype II (Georgia 2017) as well as to develop and validate a sensitive and specific ASF indirect-ELISA (iELISA) for early detection of ASF. The truncated ASF-p72 recombinant protein was successfully expressed in E. coli BL21/DE3 cells using the pET-SUMO plasmid. Bioinformatics analysis showed 100% homology among the new isolates of ASFV from genotype II. The ASF-p72-truncated protein was used to develop an iELISA, which had a high sensitivity (88%) and strong specificity (97%); the concordance index kappa was K = 0.872, indicating nearly perfect agreement compared to the WOAH confirmatory immunoperoxidase test. The validation results utilizing the reference sera panel from the OIE-ASF Reference Laboratory show the excellent detection capabilities of ASF antibodies up to a 1:1000 serum dilution. The inter-assay coefficient of variation (CV 10.4%) and intra-assay CV (2.8%) data show that the assay is precise and reproducible. This biotechnology advancement can be used to conduct future epidemiological research for ASF surveillance in ASF-free American countries.

Keywords

African swine feverAfrican swine fever virusAnimalsAntibodies, viralAntibodyCapsid protein p72, african swine fever virusCapsid proteinsCloning, molecularEasterEnzyme-linked immunosorbent assayEscherichia coliInfectionP72Recombinant proteiRecombinant proteinRecombinant proteinsSensitivity and specificitySwine

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Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Pathogens due to its progression and the good impact it has achieved in recent years, according to the agency WoS (JCR), it has become a reference in its field. In the year of publication of the work, 2025, it was in position 70/161, thus managing to position itself as a Q2 (Segundo Cuartil), in the category Microbiology. Notably, the journal is positioned en el Cuartil Q2 para la agencia Scopus (SJR) en la categoría .

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Leadership analysis of institutional authors