{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T19:12:06Z","timestamp":1771355526854,"version":"3.50.1"},"reference-count":27,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T00:00:00Z","timestamp":1771286400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004901","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de Minas Gerais","doi-asserted-by":"publisher","award":["BPD-00807-22"],"award-info":[{"award-number":["BPD-00807-22"]}],"id":[{"id":"10.13039\/501100004901","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004901","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de Minas Gerais","doi-asserted-by":"publisher","award":["APQ-05079-23"],"award-info":[{"award-number":["APQ-05079-23"]}],"id":[{"id":"10.13039\/501100004901","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["301555\/2023-0"],"award-info":[{"award-number":["301555\/2023-0"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    Serologic diagnosis of canine brucellosis caused by\n                    <jats:italic>Brucella canis<\/jats:italic>\n                    remains quite challenging since the currently available methods have considerable limitations, although the relevance and awareness about this zoonotic pathogen is increasing over the past few years. Therefore, the development of novel strategies is highly desirable. In this study,\n                    <jats:italic>in silico<\/jats:italic>\n                    analyses resulted in prediction of\n                    <jats:italic>B. canis<\/jats:italic>\n                    specific B cell epitopes, which were validated by synthesizing the corresponding peptides on a membrane followed by immunobloting with sera from dogs naturally infected with\n                    <jats:italic>B. canis<\/jats:italic>\n                    or uninfected controls as well as by excluding epitopes that cross reacted with sera from cattle and sheep infected with\n                    <jats:italic>B. abortus<\/jats:italic>\n                    and\n                    <jats:italic>B. ovis<\/jats:italic>\n                    , respectively. This approach resulted in the identification of 26 epitopes that reacted exclusively with sera from dogs infected with\n                    <jats:italic>B. canis<\/jats:italic>\n                    , from which the 15 with strongest signal in the immunoblot were selected and synthesized in soluble form for further analyses. The best 10 synthetic peptides (considering the noise to signal ratio) were used in combination as antigens for development of a\n                    <jats:italic>B. canis<\/jats:italic>\n                    specific indirect enzyme-linked immunosorbent assay (iELISA) protocol, which yielded improved specificity to differentiate from other common canine pathogens (\n                    <jats:italic>Leishmania<\/jats:italic>\n                    sp. and\n                    <jats:italic>Babesia<\/jats:italic>\n                    sp.) and an improved performance (100% specificity and 80% sensitivity) when compared to crude bacterial protein extracts used as antigen for iELISA. These selected epitopes were also incorporated in a multi-\n                    <jats:italic>B. canis<\/jats:italic>\n                    epitope protein that was expressed in\n                    <jats:italic>E. coli<\/jats:italic>\n                    and employed as antigen for detection of anti-\n                    <jats:italic>B. canis<\/jats:italic>\n                    IgG and IgM from naturally infected dogs, resulting in analytical performance similar to the iELISA protocol in which synthetic peptides were used as antigens (100% specificity and 75% sensitivity). The results clearly indicate that combination of detection of IgM and IgG may result in higher sensitivity. Therefore, this study provides novel tools for improving the accuracy of serologic diagnosis of canine brucellosis caused by\n                    <jats:italic>B. canis<\/jats:italic>\n                    .\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0342574","type":"journal-article","created":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T18:23:23Z","timestamp":1771352603000},"page":"e0342574","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of serologic diagnostic test based on in silico predicted synthetic peptides for Brucella canis in dogs"],"prefix":"10.1371","volume":"21","author":[{"given":"Monique Ferreira Silva","family":"Souza","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lucas dos Reis","family":"de Souza","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pamela Aparecida","family":"Lima","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9681-4284","authenticated-orcid":true,"given":"Tatyane Martins","family":"Cirilo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Samuel Alexandre Pimenta","family":"Carvalho","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lilian Lacerda","family":"Bueno","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jo\u00e3o Lu\u00eds","family":"Reis Cunha","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tatiane Alves da","family":"Paix\u00e3o","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ricardo Toshio","family":"Fujiwara","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4830-0470","authenticated-orcid":true,"given":"Renato Lima","family":"Santos","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2026,2,17]]},"reference":[{"issue":"1","key":"pone.0342574.ref001","doi-asserted-by":"crossref","DOI":"10.1093\/femsre\/fuaa045","article-title":"The one hundred year journey of the genus Brucella (Meyer and Shaw 1920)","volume":"45","author":"E Moreno","year":"2021","journal-title":"FEMS Microbiol Rev"},{"key":"pone.0342574.ref002","doi-asserted-by":"crossref","first-page":"594291","DOI":"10.3389\/fvets.2021.594291","article-title":"Canine Brucellosis: An Update","volume":"8","author":"RL Santos","year":"2021","journal-title":"Front Vet Sci"},{"issue":"1","key":"pone.0342574.ref003","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":"JPS Mol","year":"2020","journal-title":"J Vet Diagn Invest"},{"issue":"3","key":"pone.0342574.ref004","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1111\/tbed.12632","article-title":"Brucella canis infection in dogs from commercial breeding kennels in Brazil","volume":"64","author":"LB Keid","year":"2017","journal-title":"Transbound Emerg Dis"},{"issue":"1","key":"pone.0342574.ref005","doi-asserted-by":"crossref","first-page":"7203","DOI":"10.1038\/s41598-018-25651-x","article-title":"Tissue distribution and cell tropism of Brucella canis in naturally infected canine foetuses and neonates","volume":"8","author":"TD de Souza","year":"2018","journal-title":"Sci Rep"},{"issue":"3","key":"pone.0342574.ref006","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":"RB Hollett","year":"2006","journal-title":"Theriogenology"},{"issue":"5","key":"pone.0342574.ref007","article-title":"Cell and Tissue Tropism of Brucella spp","volume":"91","author":"TPd Carvalho","year":"2023","journal-title":"Infect Immun"},{"issue":"3","key":"pone.0342574.ref008","first-page":"117","article-title":"Laboratorial diagnosis of animal brucellosis","volume":"19","author":"JPdS Mol","year":"2012","journal-title":"Rev Bras Cienc Vet"},{"issue":"1","key":"pone.0342574.ref009","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":"LB Keid","year":"2009","journal-title":"Res Vet Sci"},{"issue":"1","key":"pone.0342574.ref010","doi-asserted-by":"crossref","first-page":"19106","DOI":"10.1038\/s41598-025-04244-5","article-title":"Preparation of a Brucella multiepitope fusion protein based on bioinformatics and its application in serological diagnosis of human brucellosis","volume":"15","author":"G Wang","year":"2025","journal-title":"Sci Rep"},{"key":"pone.0342574.ref011","doi-asserted-by":"crossref","first-page":"5301","DOI":"10.2147\/IDR.S374432","article-title":"Comparison of BP26, Omp25 and Omp31 and a Multiepitope-Based Fusion Protein in the Serological Detection of Canine Brucellosis","volume":"15","author":"M Yao","year":"2022","journal-title":"Infect Drug Resist"},{"issue":"8","key":"pone.0342574.ref012","article-title":"Paper-based ELISA diagnosis technology for human brucellosis based on a multiepitope fusion protein","volume":"15","author":"D Yin","year":"2021","journal-title":"PLoS Negl Trop Dis"},{"key":"pone.0342574.ref013","doi-asserted-by":"crossref","first-page":"708008","DOI":"10.3389\/fvets.2021.708008","article-title":"A Multi-Epitope Fusion Protein-Based p-ELISA Method for Diagnosing Bovine and Goat Brucellosis","volume":"8","author":"D Yin","year":"2021","journal-title":"Front Vet Sci"},{"key":"pone.0342574.ref014","doi-asserted-by":"crossref","first-page":"6348238","DOI":"10.1155\/jimr\/6348238","article-title":"In Silico Design and Characterization of a Multiepitope Vaccine Candidate Against Brucella canis Using a Reverse Vaccinology Approach","volume":"2025","author":"V Arriagada","year":"2025","journal-title":"J Immunol Res"},{"issue":"1","key":"pone.0342574.ref015","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1186\/s12865-025-00728-1","article-title":"Brucellosis novel multi-epitope vaccine design based on in silico analysis focusing on Brucella abortus","volume":"26","author":"H Gharazi","year":"2025","journal-title":"BMC Immunol"},{"issue":"8","key":"pone.0342574.ref016","article-title":"Encapsulated Brucella ovis Lacking a Putative ATP-Binding Cassette Transporter (\u0394abcBA) Protects against Wild Type Brucella ovis in Rams","volume":"10","author":"APC Silva","year":"2015","journal-title":"PLoS One"},{"key":"pone.0342574.ref017","doi-asserted-by":"crossref","DOI":"10.1093\/nar\/gkx346","article-title":"BepiPred-2.0: improving sequence-based B-cell epitope prediction using conformational epitopes","volume":"45","author":"MC Jespersen","year":"2017","journal-title":"Nucleic Acids Res"},{"issue":"16","key":"pone.0342574.ref018","doi-asserted-by":"crossref","first-page":"3433","DOI":"10.1093\/bioinformatics\/bti541","article-title":"IUPred: web server for the prediction of intrinsically unstructured regions of proteins based on estimated energy content","volume":"21","author":"Z Doszt\u00e1nyi","year":"2005","journal-title":"Bioinformatics"},{"issue":"20","key":"pone.0342574.ref019","doi-asserted-by":"crossref","first-page":"12503","DOI":"10.1073\/pnas.212508599","article-title":"Comparative genome analysis of the alpha -proteobacteria: relationships between plant and animal pathogens and host specificity","volume":"99","author":"RM Tsolis","year":"2002","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"8","key":"pone.0342574.ref020","article-title":"If You\u2019re Not Confused, You\u2019re Not Paying Attention: Ochrobactrum Is Not Brucella","volume":"61","author":"E Moreno","year":"2023","journal-title":"J Clin Microbiol"},{"issue":"1","key":"pone.0342574.ref021","first-page":"61","article-title":"Chronic prostatitis in a dog infected with a smooth strain of Brucella spp","volume":"14","author":"MJA R\u00fcegger","year":"2021","journal-title":"Braz J Vet Pathol"},{"issue":"3","key":"pone.0342574.ref022","article-title":"Characterization of periplasmic protein BP26 epitopes of Brucella melitensis reacting with murine monoclonal and sheep antibodies","volume":"7","author":"J Qiu","year":"2012","journal-title":"PLoS One"},{"issue":"6","key":"pone.0342574.ref023","doi-asserted-by":"crossref","first-page":"1695","DOI":"10.1590\/1678-6767","article-title":"Indirect ELISA for diagnosis of Brucella ovis infection in rams","volume":"66","author":"SA Fran\u00e7a","year":"2014","journal-title":"Arq Bras Med Vet e Zootec"},{"issue":"9","key":"pone.0342574.ref024","doi-asserted-by":"crossref","first-page":"1410","DOI":"10.1128\/CVI.00052-13","article-title":"Limitations of the BP26 protein-based indirect enzyme-linked immunosorbent assay for diagnosis of Brucellosis","volume":"20","author":"T Xin","year":"2013","journal-title":"Clin Vaccine Immunol"},{"issue":"3","key":"pone.0342574.ref025","first-page":"193","article-title":"Immunoreactivity evaluation of a new recombinant chimeric protein against Brucella in the murine model","volume":"8","author":"A Abdollahi","year":"2016","journal-title":"Iran J Microbiol"},{"issue":"4","key":"pone.0342574.ref026","first-page":"350","article-title":"Immune reactivity of sera obtained from brucellosis patients and vaccinated-rabbits to a fusion protein from Brucella melitensis","volume":"18","author":"J Amani","year":"2015","journal-title":"Iran J Basic Med Sci"},{"issue":"1","key":"pone.0342574.ref027","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1186\/1297-9716-43-29","article-title":"What have we learned from brucellosis in the mouse model?","volume":"43","author":"M-J Grill\u00f3","year":"2012","journal-title":"Vet Res"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0342574","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T18:23:32Z","timestamp":1771352612000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0342574"}},"subtitle":[],"editor":[{"given":"Odir Antonio","family":"Dellagostin","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2026,2,17]]},"references-count":27,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,2,17]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0342574","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,17]]}}}