{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T04:46:33Z","timestamp":1789620393550,"version":"build-2803163510"},"reference-count":36,"publisher":"American Society for Microbiology","issue":"7","license":[{"start":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T00:00:00Z","timestamp":1783382400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T00:00:00Z","timestamp":1783382400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2026,7,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      African swine fever (ASF) is a highly contagious and fatal hemorrhagic disease of swine. While vaccines have been licensed in Vietnam and the Philippines, their global application remains constrained by insufficient large-scale validation of safety and efficacy, rendering rapid and reliable diagnostic assays crucial for effective prevention and control. In this study, a multi-epitope antigen (MEA) was rationally designed by utilizing informatics to predict and select 15 conserved B-cell epitopes from four major African swine fever virus (ASFV) structural proteins: p30, p54, p72, and pA104R. The recombinant MEA was expressed in\n                      <jats:italic toggle=\"yes\">Escherichia coli,<\/jats:italic>\n                      and its immunogenicity was validated by Western blot and indirect immunofluorescence assays. This antigen was subsequently utilized as the basis for an indirect enzyme-linked immunosorbent assay (iELISA) whose reaction conditions were systematically optimized. The optimal cutoff value for the iELISA was determined to be 0.315 from the analysis of 100 known ASFV-negative serum samples. The assay demonstrated excellent reproducibility, with intra- and inter-assay coefficients of variation below 10%, and exhibited high specificity, with no cross-reactivity against sera positive for other common swine viruses. Notably, the developed assay exhibited high sensitivity, capable of detecting ASFV antibodies in serum samples at dilutions up to 1:6,400. Furthermore, it demonstrated superior sensitivity compared to two commercial ELISA kits in validating clinical samples from regions with circulating Genotype II strains. These findings highlight the successful development of a sensitive and specific iELISA for ASFV antibody detection, representing a valuable new tool for large-scale epidemiological surveillance and the implementation of robust control strategies for ASF.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>African swine fever (ASF) is a devastating viral disease of swine causing substantial economic losses globally. Although vaccines have recently been licensed in countries such as Vietnam and the Philippines, the global availability of a universally standardized and safe vaccine remains a significant challenge. Control efforts continue to rely heavily on rapid and accurate diagnosis. A significant limitation of current serological assays is their variable sensitivity, which can lead to missed detections and compromise surveillance programs. This study addresses this deficiency through a rational, epitope-based engineering approach. We designed a single, chimeric antigen by genetically fusing 15 conserved, immunodominant B-cell epitopes from four major African swine fever virus proteins. The resulting indirect enzyme-linked immunosorbent assay demonstrated high specificity, excellent reproducibility, and, critically, superior diagnostic sensitivity compared to two commercial kits. This work reports the development of a significantly improved serodiagnostic tool, offering enhanced reliability for large-scale epidemiological surveillance and supporting more robust control strategies against ASF.<\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.03525-25","type":"journal-article","created":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T13:02:09Z","timestamp":1779368529000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Development of an indirect ELISA based on a multi-epitope recombinant protein for antibody detection against African swine fever virus"],"prefix":"10.1128","volume":"14","author":[{"given":"Lixinjie","family":"Liu","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023b72294","id-type":"ROR","asserted-by":"publisher"}],"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary, Huazhong Agricultural University","place":["Wuhan, China"]},{"name":"Key Laboratory of Preventive Veterinary Medicine in Hubei Province, The Cooperative Innovation Center for Sustainable Pig Production","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qichao","family":"Chen","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023b72294","id-type":"ROR","asserted-by":"publisher"}],"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary, Huazhong Agricultural University","place":["Wuhan, China"]},{"name":"Key Laboratory of Preventive Veterinary Medicine in Hubei Province, The Cooperative Innovation Center for Sustainable Pig Production","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linmeng","family":"Yu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023b72294","id-type":"ROR","asserted-by":"publisher"}],"name":"National 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