{"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":1789620393305,"version":"build-2803163510"},"reference-count":18,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,3,28]],"date-time":"2026-03-28T00:00:00Z","timestamp":1774656000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"German Federal Ministry of Agriculture, Food and Regional Identity"},{"id":[{"id":"https:\/\/ror.org\/04jw21793","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>The continuous and rapid spread of African swine fever (ASF) still poses a significant threat to Eurasian wild boar and domestic pigs, leading to substantial economic losses in all associated sectors annually. In Europe, including Germany, affected wild boar populations have become an important driver and host of ASF virus (ASFV), and monitoring and surveillance is key to tailor control measures that impede viral spread. While molecular methods are used to confirm the disease and to monitor viral evolution, serology gains importance in endemically affected regions as it provides insights into disease dynamics and possible attenuation of ASFV strains. Frontline serological diagnosis is done using ELISA assays, of which several are commercialized. However, accurate performance of ELISA assays is key for correct interpretation of wild boar samples. Due to the various stages of hemolysis and decay, field samples from wild boar can be challenging for ELISA assays. To assess which indirect or competitive ELISA kit performs best when dealing with such samples, we compared the sensitivity and specificity of four commercially available ELISA kits that are licensed in Germany, as well as three unlicensed but commercially available kits. For this comparison, we used all wild boar samples submitted to the National Reference Laboratory (NRL) for ASF in years 2021 and 2022, as well as samples from domestic pigs to have a control cohort where sample quality is optimal. We observed that wild boar samples, varying in stage of hemolysis and decay, were challenging for all kits included in this study. However, samples of domestic pigs were largely interpreted correctly by ELISA, using immunoperoxidase test as verification method. Additionally, the comparability of results obtained by regional laboratories was high. Our study provides data that highlights the importance of careful kit selection, assessment of sample quality, and data interpretation for effective ASFV surveillance and control.<\/jats:p>","DOI":"10.3390\/pathogens15040360","type":"journal-article","created":{"date-parts":[[2026,3,30]],"date-time":"2026-03-30T12:11:59Z","timestamp":1774872719000},"page":"360","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Sensitivity and Specificity Assessment of Various African Swine Fever ELISA Kits for Accurate Detection of Seropositive Wild Boar"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5044-6508","authenticated-orcid":false,"given":"Virginia","family":"Friedrichs","sequence":"first","affiliation":[{"name":"Friedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4258-0651","authenticated-orcid":false,"given":"Alexander","family":"Sch\u00e4fer","sequence":"additional","affiliation":[{"name":"Friedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2717-1358","authenticated-orcid":false,"given":"Paul","family":"Deutschmann","sequence":"additional","affiliation":[{"name":"Friedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, Germany"},{"name":"Landesuntersuchungsanstalt f\u00fcr das Gesundheits- und Veterin\u00e4rwesen Sachsen (LUA), Jaegerstrasse 8\/10, 01099 Dresden, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabine","family":"Bock","sequence":"additional","affiliation":[{"name":"Landeslabor Berlin-Brandenburg (LLBB), Rudower Chaussee 39, 12489 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Hlinak","sequence":"additional","affiliation":[{"name":"Landeslabor Berlin-Brandenburg (LLBB), Rudower Chaussee 39, 12489 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wulf-Iwo","family":"Bock","sequence":"additional","affiliation":[{"name":"Landeslabor Berlin-Brandenburg (LLBB), Rudower Chaussee 39, 12489 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Moss","sequence":"additional","affiliation":[{"name":"Nieders\u00e4chsisches Landesamt f\u00fcr Verbraucherschutz und Lebensmittelsicherheit (LAVES), Stau 75, 26122 Oldenburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Beer","sequence":"additional","affiliation":[{"name":"Friedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5465-5609","authenticated-orcid":false,"given":"Sandra","family":"Blome","sequence":"additional","affiliation":[{"name":"Friedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Sauter-Louis, C., Conraths, F.J., Probst, C., Blohm, U., Schulz, K., Sehl, J., Fischer, M., Forth, J.H., Zani, L., and Depner, K. (2021). African Swine Fever in Wild Boar in Europe-A Review. Viruses, 13.","DOI":"10.3390\/v13091717"},{"key":"ref_2","unstructured":"Beltran-Alcrudo, D., Arias, M., Gallardo, C., Kramer, S., and Penrith, M.L. (2017). African Swine Fever: Detection and Diagnosis\u2014A Manual for Veterinarians, Food and Agriculture Organization of the United Nations (FAO). FAO Animal Production and Health Manual No. 19."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"197676","DOI":"10.1016\/j.virusres.2019.197676","article-title":"African swine fever (ASF) diagnosis, an essential tool in the epidemiological investigation","volume":"271","author":"Gallardo","year":"2019","journal-title":"Virus Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1146\/annurev-animal-021419-083741","article-title":"African Swine Fever Epidemiology and Control","volume":"8","author":"Dixon","year":"2020","journal-title":"Annu. Rev. Anim. Biosci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1186\/s13567-015-0289-6","article-title":"How to survey classical swine fever in wild boar (Sus scrofa) after the completion of oral vaccination? Chasing away the ghost of infection at different spatial scales","volume":"47","author":"Saubusse","year":"2016","journal-title":"Vet. Res."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Probst, C., Gethmann, J., Amendt, J., Lutz, L., Teifke, J.P., and Conraths, F.J. (2020). Estimating the Postmortem Interval of Wild Boar Carcasses. Vet. Sci., 7.","DOI":"10.3390\/vetsci7010006"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e12305","DOI":"10.1002\/2688-8319.12305","article-title":"Season, decay stage, habitat, temperature and carrion beetles allow estimating the post-mortem interval of wild boar carcasses","volume":"5","author":"Rietz","year":"2024","journal-title":"Ecol. Solut. Evid."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"4151","DOI":"10.1038\/s41598-022-08142-y","article-title":"Chronological expression and distribution of African swine fever virus p30 and p72 proteins in experimentally infected pigs","volume":"12","author":"Oh","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"9959847","DOI":"10.1155\/2023\/9959847","article-title":"Dynamics of Serological and Mucosal Antibody Responses against African Swine Fever Viruses in Experimentally Infected Pigs","volume":"2023","author":"Xie","year":"2023","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1665","DOI":"10.1111\/tbed.12717","article-title":"Diagnostic specificity of the African swine fever virus antibody detection enzyme-linked immunosorbent assay in feral and domestic pigs in the United States","volume":"64","author":"Bergeron","year":"2017","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"469","DOI":"10.4103\/0301-4738.71673","article-title":"Understanding the relevance of sample size calculation","volume":"58","author":"Nayak","year":"2010","journal-title":"Indian J. Ophthalmol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1590\/2176-9451.19.4.027-029.ebo","article-title":"How sample size influences research outcomes","volume":"19","author":"Faber","year":"2014","journal-title":"Dent. Press J. Orthod."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Gimenez-Lirola, L.G., Mur, L., Rivera, B., Mogler, M., Sun, Y., Lizano, S., Goodell, C., Harris, D.L., Rowland, R.R., and Gallardo, C. (2016). Detection of African Swine Fever Virus Antibodies in Serum and Oral Fluid Specimens Using a Recombinant Protein 30 (p30) Dual Matrix Indirect ELISA. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0161230"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.molimm.2025.07.009","article-title":"Preparation of African swine fever virus (ASFV) p30 monoclonal antibodies and identification of novel antigenic epitopes","volume":"185","author":"Zhang","year":"2025","journal-title":"Mol. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Pikalo, J., Carrau, T., Deutschmann, P., Fischer, M., Schlottau, K., Beer, M., and Blome, S. (2022). Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection-Comparison of Twelve Kits to an OIE-Recommended Method. Viruses, 14.","DOI":"10.3390\/v14020220"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e3231","DOI":"10.1111\/tbed.14491","article-title":"Comparison of the sensitivity, specificity, correlation and inter-assay agreement of eight diagnostic in vitro assays for the detection of African swine fever virus","volume":"69","author":"Auer","year":"2022","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"23958","DOI":"10.1038\/s41598-024-75793-4","article-title":"Evaluation and comparison of performances of six commercial NSP ELISA assays for foot and mouth disease virus in Thailand","volume":"14","author":"Seeyo","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Aguilar-Vargas, F., Carvajal, F., Guzm\u00e1n, M., and Le\u00f3n, B. (2023). Comparison of three commercial enzyme-linked immunosorbent assays for the diagnosis of Classical Swine Fever. Arch. Vet. Sci., 28.","DOI":"10.5380\/avs.v28i2.90894"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/15\/4\/360\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T16:02:44Z","timestamp":1775059364000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/15\/4\/360"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,28]]},"references-count":18,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["pathogens15040360"],"URL":"https:\/\/doi.org\/10.3390\/pathogens15040360","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,28]]}}}