{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T19:28:24Z","timestamp":1792092504944,"version":"4.4.0"},"reference-count":52,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2023,8,26]],"date-time":"2023-08-26T00:00:00Z","timestamp":1693008000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Italian Ministry of Health","award":["IZSAM 02\/20 RC"],"award-info":[{"award-number":["IZSAM 02\/20 RC"]}]},{"name":"Italian Ministry of Health","award":["IZSAM 02\/21 RC"],"award-info":[{"award-number":["IZSAM 02\/21 RC"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Canine brucellosis caused by Brucella canis, is an infectious disease affecting dogs and wild Canidae. Clinical diagnosis is challenging, and laboratory testing is crucial for a definitive diagnosis. Various serological methods have been described, but their accuracy is uncertain due to limited validation studies. The present study aimed to evaluate the performances of three serological tests for the diagnosis of B. canis in comparison with bacterial isolation (gold standard), in order to establish a protocol for the serological diagnosis of canine brucellosis. A panel of sera from naturally infected dogs (n = 61), from which B. canis was isolated, and uninfected dogs (n = 143), negative for B. canis isolation, were tested using microplate serum agglutination (mSAT), complement fixation performed using the Brucella ovis antigen (B. ovis-CFT), and a commercial immunofluorescence assay (IFAT). The sensitivity and specificity of the three serological methods were, respectively, the following: 96.7% (95% CI 88.8\u201398.7%) and 92.3 (95% CI 86.7\u201395.1%) for mSAT; 96.7% (95% CI 88.8\u201398.7%) and 96.5 (95% CI 92.1\u201398.2%) for B. ovis-CFT; 98.4% (95% CI 91.3\u201399.4%) and 99.3 (95% CI 96.2\u201399.8%) for IFAT. The use in of the three methods in parallel, combined with bacterial isolation and molecular methods, could improve the diagnosis of the infection in dogs.<\/jats:p>","DOI":"10.3390\/microorganisms11092162","type":"journal-article","created":{"date-parts":[[2023,8,28]],"date-time":"2023-08-28T02:23:25Z","timestamp":1693189405000},"page":"2162","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Evaluation of Three Serological Tests for Diagnosis of Canine Brucellosis"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7477-404X","authenticated-orcid":false,"given":"Fabrizia","family":"Perletta","sequence":"first","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2694-2835","authenticated-orcid":false,"given":"Chiara","family":"Di Pancrazio","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diamante","family":"Rodomonti","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tiziana","family":"Di Febo","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0866-0869","authenticated-orcid":false,"given":"Mirella","family":"Luciani","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0852-2797","authenticated-orcid":false,"given":"Ivanka Marinova","family":"Krasteva","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1609-6815","authenticated-orcid":false,"given":"Marta","family":"Maggetti","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca","family":"Profeta","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7456-0073","authenticated-orcid":false,"given":"Romolo","family":"Salini","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9811-699X","authenticated-orcid":false,"given":"Fabrizio","family":"De Massis","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2618-6192","authenticated-orcid":false,"given":"Flavio","family":"Sacchini","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6614-9989","authenticated-orcid":false,"given":"Manuela","family":"Tittarelli","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e del Molise, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,8,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1292\/jvms.70.707","article-title":"Evaluation of a microplate agglutination test (MAT) for serological diagnosis of canine brucellosis","volume":"70","author":"Kimura","year":"2008","journal-title":"J. Vet. Med. Sci."},{"key":"ref_2","unstructured":"Nielsen, K., and Duncan, J.R. (1990). Animal Brucellosis, CRC Press."},{"key":"ref_3","unstructured":"Greene, C.E. (2012). Infectious Diseases of Dog and Cat, Elsevier Saunders. [4th ed.]."},{"key":"ref_4","first-page":"1589","article-title":"Semen examination in dogs with canine brucellosis","volume":"40","author":"George","year":"1979","journal-title":"Am. J. Vet. Res."},{"key":"ref_5","first-page":"371","article-title":"Problems in the Serodiagnosis of Canine Brucellosis\u2014Dog-Responses to Cell-Wall and Internal Antigens of Brucella-Canis","volume":"56","author":"Carmichael","year":"1984","journal-title":"Dev. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1043610","DOI":"10.3389\/fvets.2022.1043610","article-title":"Brucella canis discospondylitis in 33 dogs","volume":"9","author":"Long","year":"2022","journal-title":"Front. Vet. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1233","DOI":"10.1007\/s00580-013-1767-8","article-title":"Brucella canis-induced anterior uveitis in companion dogs in Ahvaz, Iran","volume":"23","author":"Mosallanejad","year":"2014","journal-title":"Comp. Clin. Pathol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1292\/jvms1939.39.181","article-title":"Ocular lesions in experimental canine brucellosis","volume":"39","author":"Saeguesa","year":"1977","journal-title":"Jpn. J. Vet. Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"4861","DOI":"10.1128\/IAI.00995-15","article-title":"Brucella canis Is an Intracellular Pathogen That Induces a Lower Proinflammatory Response than Smooth Zoonotic Counterparts","volume":"83","author":"Medina","year":"2015","journal-title":"Infect. Immun."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1016\/j.rvsc.2007.02.006","article-title":"Diagnosis of canine brucellosis by ELISA using an antigen obtained from wild Brucella canis","volume":"83","year":"2007","journal-title":"Res. Vet. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/j.anireprosci.2004.05.005","article-title":"Canine brucellosis","volume":"82\u201383","author":"Wanke","year":"2004","journal-title":"Anim. Reprod. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1401","DOI":"10.3201\/eid2408.171171","article-title":"Brucellosis in Dogs and Public Health Risk","volume":"24","author":"Hensel","year":"2018","journal-title":"Emerg. Infect. Dis."},{"key":"ref_13","first-page":"779","article-title":"Seroprevalence of Brucella canis infection of dogs in two provinces in Turkey","volume":"29","author":"Oncel","year":"2005","journal-title":"Turk. J. Vet. Anim. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"151","DOI":"10.3389\/fvets.2019.00151","article-title":"Canine Brucellosis: Insights Into the Epidemiologic Situation in Europe","volume":"6","author":"Buhmann","year":"2019","journal-title":"Front. Vet. Sci."},{"key":"ref_15","first-page":"251","article-title":"Outbreak of a Brucella canis infection in a beagle colony in West Germany","volume":"31","year":"1976","journal-title":"Dev. Biol. Stand."},{"key":"ref_16","first-page":"368","article-title":"Canine brucellosis\u2014A case report","volume":"116","author":"Nockler","year":"2003","journal-title":"Berl. Munch. Tierarztl. Wochenschr."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1177\/104063871102300127","article-title":"Detection of Brucella canis-induced reproductive diseases in a kennel","volume":"23","author":"Gyuranecz","year":"2011","journal-title":"J. Vet. Diagn. Invest."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1016\/j.vetmic.2014.10.015","article-title":"Brucellosis outbreak in a Swedish kennel in 2013: Determination of genetic markers for source tracing","volume":"174","author":"Kaden","year":"2014","journal-title":"Vet. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"743","DOI":"10.17236\/sat00190","article-title":"Brucella canis infection in a young dog with epididymitis and orchitis","volume":"160","author":"Egloff","year":"2018","journal-title":"Schweiz. Arch. Tierheilkd."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1136\/vr.106.5.102","article-title":"Serological Evidence for the Presence of Brucella canis Infection in Dogs in Britain","volume":"106","author":"Taylor","year":"1980","journal-title":"Vet. Rec."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1136\/vr.j3004","article-title":"Second UK isolation of Brucella canis","volume":"180","author":"Whatmore","year":"2017","journal-title":"Vet. Rec."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1136\/vr.j1811","article-title":"Brucella canis in a dog in the UK","volume":"180","author":"Morgan","year":"2017","journal-title":"Vet. Rec."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"665","DOI":"10.3906\/vet-1803-54","article-title":"The first-time isolation of Brucella canis from two aborted bitches in a kennel in Turkey","volume":"42","author":"Saytekin","year":"2018","journal-title":"Turk. J. Vet. Anim. Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"5","DOI":"10.36016\/JVMBBS-2020-6-4-1","article-title":"First report of Canine brucellosis in Ukraine: Pathogen isolation and characterization","volume":"6","author":"Bolotin","year":"2020","journal-title":"J. Vet. Med. Biotechnol. Biosaf."},{"key":"ref_25","first-page":"215","article-title":"First Isolation of Brucella canis from a breeding kennel in Italy","volume":"57","author":"Sacchini","year":"2021","journal-title":"Vet. Ital."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1783","DOI":"10.3201\/eid2707.201238","article-title":"Transboundary Spread of Brucella canis through Import of Infected Dogs, the Netherlands, November 2016\u2013December 2018","volume":"27","author":"Engelsma","year":"2021","journal-title":"Emerg. Infect. Dis."},{"key":"ref_27","unstructured":"Girault, G., Ferreira Vicente, A., Ribeiro, M., Perrot, L., Petot-Bottin, F., Freddi, L., Djokic, V., and Ponsart, C. (October, January 29). Canine Brucellosis in France due to Brucella canis: An emerging disease?. Proceedings of the Brucellosis 2022 International Research Conference Giulianova, Teramo, Italy."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1111\/zph.12163","article-title":"Human Brucella canis Infection and Subsequent Laboratory Exposures Associated with a Puppy, New York City, 2012","volume":"62","author":"Dentinger","year":"2015","journal-title":"Zoonoses Public Health"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e62776","DOI":"10.5812\/archcid.62776","article-title":"Human Brucellosis Caused by Brucella canis: A Rare Case Report","volume":"13","author":"Majzoobi","year":"2018","journal-title":"Arch. Clin. Infect. Dis."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2250","DOI":"10.1093\/cid\/ciac425","article-title":"First Case of Human Brucella canis Infection in the Netherlands","volume":"75","author":"Kolwijck","year":"2022","journal-title":"Clin. Infect. Dis."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"457","DOI":"10.1099\/jmm.0.45927-0","article-title":"Diagnosis of human brucellosis caused by Brucella canis","volume":"54","author":"Lucero","year":"2005","journal-title":"J. Med. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1017\/S0950268809990525","article-title":"Human Brucella canis outbreak linked to infection in dogs","volume":"138","author":"Lucero","year":"2010","journal-title":"Epidemiol. Infect."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2489","DOI":"10.3201\/eid2709.204701","article-title":"Laboratory Exposures from an Unsuspected Case of Human Infection with Brucella canis","volume":"27","author":"Roman","year":"2021","journal-title":"Emerg. Infect. Dis."},{"key":"ref_34","unstructured":"WOAH (2022). Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, WOAH. Available online: https:\/\/www.woah.org\/fileadmin\/Home\/fr\/Health_standards\/tahm\/3.01.04_BRUCELLOSIS.pdf."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"959328","DOI":"10.3389\/fimmu.2022.959328","article-title":"Primary and memory immune responses against rough Brucella canis are less robust compared to smooth B. abortus and B. melitensis following intratracheal infection in mice","volume":"13","author":"Stranahan","year":"2022","journal-title":"Front. Immunol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1045","DOI":"10.1189\/jlb.0103015","article-title":"Smooth and rough lipopolysaccharide phenotypes of Brucella induce different intracellular trafficking and cytokine\/chemokine release in human monocytes","volume":"74","author":"Rittig","year":"2003","journal-title":"J. Leukoc. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/j.vetmic.2004.10.020","article-title":"Use of Brucella canis antigen for detection of ovine serum antibodies against Brucella ovis","volume":"105","author":"Lopez","year":"2005","journal-title":"Vet. Microbiol."},{"key":"ref_38","first-page":"5","article-title":"Canine brucellosis due to Brucella canis: Description of the disease and control measures","volume":"58","author":"Sacchini","year":"2022","journal-title":"Vet. Ital."},{"key":"ref_39","unstructured":"Alton, G.G., Jones, L.M., Angus, R.D., and Verger, J.M. (1988). Techniques for the Brucellosis Laboratory, INRA."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1016\/j.theriogenology.2006.04.011","article-title":"Canine brucellosis: Outbreaks and compliance","volume":"66","author":"Hollett","year":"2006","journal-title":"Theriogenology"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1186\/1751-0147-54-18","article-title":"The first case of Brucella canis in Sweden: Background, case report and recommendations from a northern European perspective","volume":"54","author":"Holst","year":"2012","journal-title":"Acta Vet. Scand."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1177\/1040638719891083","article-title":"Diagnosis of canine brucellosis: Comparison of various serologic tests and PCR","volume":"32","author":"Mol","year":"2020","journal-title":"J. Vet. Diagn. Investig."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"594291","DOI":"10.3389\/fvets.2021.594291","article-title":"Canine Brucellosis: An Update","volume":"8","author":"Santos","year":"2021","journal-title":"Front. Vet. Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"516","DOI":"10.7883\/yoken.64.516","article-title":"A Serological Diagnostic Survey for Brucella canis Infection in Turkish Patients with Brucellosis-Like Symptoms","volume":"64","author":"Sayan","year":"2011","journal-title":"Jpn. J. Infect. Dis."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2166","DOI":"10.1128\/jcm.33.8.2166-2170.1995","article-title":"Comparative-Evaluation of Microagglutination Test and Serum Agglutination-Test as Supplementary Diagnostic Methods for Brucellosis","volume":"33","author":"Baum","year":"1995","journal-title":"J. Clin. Microbiol."},{"key":"ref_46","unstructured":"Alton, G.G., Jones, L.M., Angus, R.D., and Verger, J.M. (1988). Techniques for the Brucellosis Laboratory, INRA."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"986","DOI":"10.1093\/clinchem\/42.6.986","article-title":"Presentation of receiver-operating characteristic (ROC) plots","volume":"42","author":"Schoonjans","year":"1996","journal-title":"Clin. Chem."},{"key":"ref_48","unstructured":"Sivia, D.S. (1996). Data Analysis: A Bayesian Tutorial, Oxford University Press."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1099\/0022-1317-50-8-663","article-title":"Evaluation of three methods to measure anti-Brucella IgM antibodies and interference of IgA in the interpretation of mercaptan-based tests","volume":"50","author":"Marrodan","year":"2001","journal-title":"J. Med. Microbiol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.rvsc.2008.05.012","article-title":"Comparison of agar gel immunodiffusion test, rapid slide agglutination test, microbiological culture and PCR for the diagnosis of canine brucellosis","volume":"86","author":"Keid","year":"2009","journal-title":"Res. Vet. Sci."},{"key":"ref_51","first-page":"74","article-title":"Brucella canis: An update on research and clinical management","volume":"59","author":"Cosford","year":"2018","journal-title":"Can. Vet. J.-Rev. Vet. Can."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"951","DOI":"10.1007\/s11259-006-0109-6","article-title":"Diagnosis of canine brucellosis: Comparison between serological and microbiological tests and a PCR based on primers to 16S-23S rDNA interspacer","volume":"31","author":"Keid","year":"2007","journal-title":"Vet. Res. Commun."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/11\/9\/2162\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:39:39Z","timestamp":1760128779000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/11\/9\/2162"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,8,26]]},"references-count":52,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2023,9]]}},"alternative-id":["microorganisms11092162"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms11092162","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,8,26]]}}}