{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T02:17:41Z","timestamp":1760235461382,"version":"build-2065373602"},"reference-count":36,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2021,8,19]],"date-time":"2021-08-19T00:00:00Z","timestamp":1629331200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000072","name":"National Institute of Dental and Craniofacial Research","doi-asserted-by":"publisher","award":["Intramural Research Program"],"award-info":[{"award-number":["Intramural Research Program"]}],"id":[{"id":"10.13039\/100000072","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Government of Japan for the IAEA Peaceful Uses Initiative","award":["Project \u201cDetection of emerging and re-emerging animal and zoonotic pathogens at the animal-human interface"],"award-info":[{"award-number":["Project \u201cDetection of emerging and re-emerging animal and zoonotic pathogens at the animal-human interface"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The recent emergence of SARS-CoV-2 in humans from a yet unidentified animal reservoir and the capacity of the virus to naturally infect pets, farmed animals and potentially wild animals has highlighted the need for serological surveillance tools. In this study, the luciferase immunoprecipitation systems (LIPS), employing the spike (S) and nucleocapsid proteins (N) of SARS-CoV-2, was used to examine the suitability of the assay for antibody detection in different animal species. Sera from SARS-CoV-2 naturally-infected mink (n = 77), SARS-CoV-2 experimentally-infected ferrets, fruit bats and hamsters and a rabbit vaccinated with a purified spike protein were examined for antibodies using the SARS-CoV-2 N and\/or S proteins. From comparison with the known neutralization status of the serum samples, statistical analyses including calculation of the Spearman rank-order-correlation coefficient and Cohen\u2019s kappa agreement were used to interpret the antibody results and diagnostic performance. The LIPS immunoassay robustly detected the presence of viral antibodies in naturally infected SARS-CoV-2 mink, experimentally infected ferrets, fruit bats and hamsters as well as in an immunized rabbit. For the SARS-CoV-2-LIPS-S assay, there was a good level of discrimination between the positive and negative samples for each of the five species tested with 100% agreement with the virus neutralization results. In contrast, the SARS-CoV-2-LIPS-N assay did not consistently differentiate between SARS-CoV-2 positive and negative sera. This study demonstrates the suitability of the SARS-CoV-2-LIPS-S assay for the sero-surveillance of SARS-CoV-2 infection in a range of animal species.<\/jats:p>","DOI":"10.3390\/v13081649","type":"journal-article","created":{"date-parts":[[2021,8,19]],"date-time":"2021-08-19T09:58:06Z","timestamp":1629367086000},"page":"1649","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Serological Detection of SARS-CoV-2 Antibodies in Naturally-Infected Mink and Other Experimentally-Infected Animals"],"prefix":"10.3390","volume":"13","author":[{"given":"Francisco J.","family":"Berguido","sequence":"first","affiliation":[{"name":"Joint FAO\/IAEA Centre for Nuclear Applications in Food and Agriculture, Animal Production and Health Laboratory, Department of Nuclear Sciences and Applications, International Atomic Energy Agency Vienna International Centre, P.O. Box 100, 1400 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1717-048X","authenticated-orcid":false,"given":"Peter D.","family":"Burbelo","sequence":"additional","affiliation":[{"name":"National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alessio","family":"Bortolami","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Animal Models, Division of Comparative Biomedical Sciences, Istituto Zooprofilattico Sperimentale delle Venezie, 35020 Legnaro, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesco","family":"Bonfante","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Animal Models, Division of Comparative Biomedical Sciences, Istituto Zooprofilattico Sperimentale delle Venezie, 35020 Legnaro, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8071-0827","authenticated-orcid":false,"given":"Kerstin","family":"Wernike","sequence":"additional","affiliation":[{"name":"Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, S\u00fcdufer 10, 17493 Greifswald, Insel Riems, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4552-031X","authenticated-orcid":false,"given":"Donata","family":"Hoffmann","sequence":"additional","affiliation":[{"name":"Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, S\u00fcdufer 10, 17493 Greifswald, Insel Riems, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7613-9592","authenticated-orcid":false,"given":"Anne","family":"Balkema-Buschmann","sequence":"additional","affiliation":[{"name":"Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, S\u00fcdufer 10, 17493 Greifswald, Insel Riems, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martin","family":"Beer","sequence":"additional","affiliation":[{"name":"Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, S\u00fcdufer 10, 17493 Greifswald, Insel Riems, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2776-169X","authenticated-orcid":false,"given":"William G.","family":"Dundon","sequence":"additional","affiliation":[{"name":"Joint FAO\/IAEA Centre for Nuclear Applications in Food and Agriculture, Animal Production and Health Laboratory, Department of Nuclear Sciences and Applications, International Atomic Energy Agency Vienna International Centre, P.O. Box 100, 1400 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1899-7614","authenticated-orcid":false,"given":"Charles E.","family":"Lamien","sequence":"additional","affiliation":[{"name":"Joint FAO\/IAEA Centre for Nuclear Applications in Food and Agriculture, Animal Production and Health Laboratory, Department of Nuclear Sciences and Applications, International Atomic Energy Agency Vienna International Centre, P.O. Box 100, 1400 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giovanni","family":"Cattoli","sequence":"additional","affiliation":[{"name":"Joint FAO\/IAEA Centre for Nuclear Applications in Food and Agriculture, Animal Production and Health Laboratory, Department of Nuclear Sciences and Applications, International Atomic Energy Agency Vienna International Centre, P.O. Box 100, 1400 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,8,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/S0140-6736(20)30251-8","article-title":"Genomic characterisation and epidemiology of 2019 novel coronavirus: Implications for virus origins and receptor binding","volume":"395","author":"Lu","year":"2020","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s11259-021-09787-2","article-title":"Bats, pangolins, minks and other animals\u2014villains or victims of SARS-CoV-2?","volume":"45","author":"Lopes","year":"2021","journal-title":"Vet. Res. Commun."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/s42522-021-00039-6","article-title":"Assessing the risks of SARS-CoV-2 in wildlife","volume":"3","author":"Delahay","year":"2021","journal-title":"One Health Outlook"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Logeot, M., Mauroy, A., Thiry, E., De Regge, N., Vervaeke, M., Beck, O., De Waele, V., and Van den Berg, T. (2021). Risk assessment of SARS-CoV-2 infection in free-ranging wild animals in Belgium. Transbound. Emerg. Dis., in press.","DOI":"10.1111\/tbed.14131"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Aguil\u00f3-Gisbert, J., Padilla-Blanco, M., Lizana, V., Maiques, E., Mu\u00f1oz-Baquero, M., Chillida-Mart\u00ednez, E., Cardells, J., and Rubio-Guerri, C. (2021). First description of SARS-CoV-2 infection in two feral American mink (Neovison vison) caught in the wild. Animals, 11.","DOI":"10.20944\/preprints202103.0647.v1"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1051\/vetres:2005053","article-title":"Genetic evolution of canine coronavirus and recent advances in prophylaxis","volume":"37","author":"Pratelli","year":"2006","journal-title":"Vet. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.tim.2016.09.001","article-title":"Molecular evolution of human coronavirus genomes","volume":"25","author":"Forni","year":"2017","journal-title":"Trends Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.3201\/eid2704.203945","article-title":"Animal reservoirs and hosts for emerging alphacoronaviruses and betacoronaviruses","volume":"27","author":"Ghai","year":"2021","journal-title":"Emerg. Infect. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Vlasova, A.N., Diaz, A., Damtie, D., Xiu, L., Toh, T.H., Lee, J.S., Saif, L.J., and Gray, G.C. (2021). Novel canine coronavirus isolated from a hospitalized pneumonia patient, East Malaysia. Clin. Infect. Dis., in press.","DOI":"10.1093\/cid\/ciab456"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Palmer, M.V., Martins, M., Falkenberg, S., Buckley, A., Caserta, L.C., Mitchell, P.K., Cassmann, E.D., Rollins, A., Zylich, N.C., and Renshaw, R.W. (2021). Susceptibility of white-tailed deer (Odocoileus virginianus) to SARS-CoV-2. J. Virol., in press.","DOI":"10.1101\/2021.01.13.426628"},{"key":"ref_11","first-page":"2001005","article-title":"SARS-CoV-2 infection in farmed minks, the Netherlands, April and May 2020","volume":"23","author":"Oreshkova","year":"2020","journal-title":"Euro. Surveill."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"European Food Safety Authority and European Centre for Disease Prevention and Control, Boklund, A., Gort\u00e1zar, C., Pasquali, P., Roberts, H., Nielsen, S.S., Stahl, K., Stegeman, A., Baldinelli, F., and Broglia, A. (2021). Monitoring of SARS-CoV-2 infection in mustelids. EFSA J., 19, e06459.","DOI":"10.2903\/j.efsa.2021.6459"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.2478\/jvetres-2021-0017","article-title":"Mink SARS-CoV-2 infection in Poland\u2014Short communication","volume":"65","author":"Smreczak","year":"2021","journal-title":"J. Vet. Res."},{"key":"ref_14","unstructured":"(2021, August 05). GLEWS (Joint FAO\u2013OIE\u2013WHO Global Early Warning System) 2021 SARS-CoV-2 in Animals Used for Fur Farming. Available online: WHO-2019-nCoV-fur-farming-risk-assessment-2021.1-eng.pdf."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"103101","DOI":"10.1016\/j.ebiom.2020.103101","article-title":"Analytical and clinical performances of five immunoassays for the detection of SARS-CoV-2 antibodies in comparison with neutralization activity","volume":"62","author":"Padoan","year":"2020","journal-title":"EBioMedicine"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"104512","DOI":"10.1016\/j.jcv.2020.104512","article-title":"Performance of six SARS-CoV-2 immunoassays in comparison with microneutralisation","volume":"129","author":"Kuivanen","year":"2020","journal-title":"J. Clin. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"610688","DOI":"10.3389\/fimmu.2020.610688","article-title":"Humoral responses and serological assays in SARS-CoV-2 Infections","volume":"11","author":"Galipeau","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_18","first-page":"m2516","article-title":"Diagnostic accuracy of serological tests for covid-19: Systematic review and meta-analysis","volume":"370","author":"Tavaziva","year":"2020","journal-title":"BMJ"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1093\/infdis\/jiaa273","article-title":"Sensitivity in detection of antibodies to nucleocapsid and spike proteins of severe acute respiratory syndrome coronavirus 2 in patients with coronavirus disease 2019","volume":"222","author":"Burbelo","year":"2020","journal-title":"J. Infect. Dis."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1234","DOI":"10.1002\/eji.202048715","article-title":"LIPS method for the detection of SARS-CoV-2 antibodies to spike and nucleocapsid proteins","volume":"50","author":"Remm","year":"2020","journal-title":"Eur. J. Immunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"6366","DOI":"10.1172\/JCI142804","article-title":"COVID-19 survival associates with the immunoglobulin response to the SARS-CoV-2 spike receptor binding domain","volume":"130","author":"Secchi","year":"2020","journal-title":"J. Clin. Investig."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Boadella, M., and Gort\u00e1zar, C. (2011). Effect of haemolysis and repeated freeze-thawing cycles on wild boar serum antibody testing by ELISA. BMC Res. Notes, 4.","DOI":"10.1186\/1756-0500-4-498"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.jviromet.2015.10.008","article-title":"Specific detection of peste des petits ruminants virus antibodies in sheep and goat sera by the luciferase immunoprecipitation system","volume":"227","author":"Berguido","year":"2016","journal-title":"J. Virol. Methods"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1038\/s41586-021-03361-1","article-title":"SARS-CoV-2 spike D614G change enhances replication and transmission","volume":"592","author":"Zhou","year":"2021","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Wernike, K., Aebischer, A., Michelitsch, A., Hoffmann, D., Freuling, C., Balkema-Buschmann, A., Graaf, A., M\u00fcller, T., Osterrieder, N., and Rissmann, M. (2020). Multi-species ELISA for the detection of antibodies against SARS-CoV-2 in animals. Transbound. Emerg. Dis., in press.","DOI":"10.1101\/2020.08.26.266825"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e218","DOI":"10.1016\/S2666-5247(20)30089-6","article-title":"SARS-CoV-2 in fruit bats; ferrets, pigs, and chickens: An experimental transmission study","volume":"1","author":"Schlottau","year":"2020","journal-title":"Lancet Microbe"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Calvet, G.A., Pereira, S.A., Ogrzewalska, M., Pauvolid-Corr\u00eaa, A., Resende, P.C., Tassinari, W.S., Costa, A.P., Keidel, L.O., da Rocha, A.S.B., and da Silva, M.F.B. (2021). Investigation of SARS-CoV-2 infection in dogs and cats of humans diagnosed with COVID-19 in Rio de Janeiro, Brazil. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0250853"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2982","DOI":"10.3201\/eid2612.203733","article-title":"Susceptibility of raccoon dogs for experimental SARS-CoV-2 infection","volume":"26","author":"Freuling","year":"2020","journal-title":"Emerg. Infect. Dis."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1193","DOI":"10.3201\/eid2704.204945","article-title":"Experimental SARS-CoV-2 infection of bank voles","volume":"27","author":"Ulrich","year":"2021","journal-title":"Emerg. Infect. Dis."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Zhao, Y., Yang, Y., Gao, J., Huang, K., Hu, C., Hui, X., He, X., Li, C., Gong, W., and Lv, C. (2021). A serological survey of severe acute respiratory syndrome coronavirus 2 in dogs in Wuhan. Transbound. Emerg. Dis., in press.","DOI":"10.1111\/tbed.14024"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"100164","DOI":"10.1016\/j.onehlt.2020.100164","article-title":"Absence of SARS-CoV-2 infection in cats and dogs in close contact with a cluster of COVID-19 patients in a veterinary campus","volume":"10","author":"Temmam","year":"2020","journal-title":"One Health"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e01828-20","DOI":"10.1128\/JVI.01828-20","article-title":"Changes in SARS-CoV-2 spike versus nucleoprotein antibody responses impact the estimates of infections in population-based seroprevalence studies","volume":"95","author":"Fenwick","year":"2021","journal-title":"J. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1038\/s41467-020-20439-y","article-title":"Dose-dependent response to infection with SARS-CoV-2 in the ferret model and evidence of protective immunity","volume":"12","author":"Ryan","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Shuai, L., Zhong, G., Yuan, Q., Wen, Z., Wang, C., He, X., Liu, R., Wang, J., Zhao, Q., and Liu, Y. (2021). Replication, pathogenicity, and transmission of SARS-CoV-2 in minks. Natl. Sci. Rev., in press.","DOI":"10.1093\/nsr\/nwaa291"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"704","DOI":"10.1016\/j.chom.2020.03.023","article-title":"Infection and rapid transmission of SARS-CoV-2 in ferrets","volume":"27","author":"Kim","year":"2020","journal-title":"Cell Host Microbe"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"834","DOI":"10.1038\/s41586-020-2342-5","article-title":"Pathogenesis and transmission of SARS-CoV-2 in golden hamsters","volume":"583","author":"Sia","year":"2020","journal-title":"Nature"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/8\/1649\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:47:23Z","timestamp":1760165243000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/8\/1649"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,8,19]]},"references-count":36,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2021,8]]}},"alternative-id":["v13081649"],"URL":"https:\/\/doi.org\/10.3390\/v13081649","relation":{},"ISSN":["1999-4915"],"issn-type":[{"type":"electronic","value":"1999-4915"}],"subject":[],"published":{"date-parts":[[2021,8,19]]}}}