{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,13]],"date-time":"2025-12-13T23:09:27Z","timestamp":1765667367974,"version":"3.41.2"},"reference-count":72,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,10,22]],"date-time":"2021-10-22T00:00:00Z","timestamp":1634860800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Bildung und Forschung","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p><jats:italic>Brucella melitensis<\/jats:italic> Rev.1 is a live attenuated vaccine strain that is widely used to control brucellosis in small ruminants. For successful surveillance and control programs, rapid identification and characterization of <jats:italic>Brucella<\/jats:italic> isolates and reliable differentiation of vaccinated and naturally infected animals are essential prerequisites. Although MALDI-TOF MS is increasingly applied in clinical microbiology laboratories for the diagnosis of brucellosis, species or even strain differentiation by this method remains a challenge. To detect biomarkers, which enable to distinguish the <jats:italic>B. melitensis<\/jats:italic> Rev.1 vaccine strain from <jats:italic>B. melitensis<\/jats:italic> field isolates, we initially searched for unique marker proteins by <jats:italic>in silico<\/jats:italic> comparison of the <jats:italic>B. melitensis<\/jats:italic> Rev.1 and 16M proteomes. We found 113 protein sequences of <jats:italic>B. melitensis<\/jats:italic> 16M that revealed a homologous sequence in the <jats:italic>B. melitensis<\/jats:italic> Rev.1 annotation and 17 of these sequences yielded potential biomarker pairs. MALDI-TOF MS spectra of 18 <jats:italic>B. melitensis<\/jats:italic> Rev.1 vaccine and 183 Israeli <jats:italic>B. melitensis<\/jats:italic> field isolates were subsequently analyzed to validate the identified marker candidates. This approach detected two genus-wide unique biomarkers with properties most similar to the ribosomal proteins L24 and S12. These two proteins clearly discriminated <jats:italic>B. melitensis<\/jats:italic> Rev.1 from the closely related <jats:italic>B. melitensis<\/jats:italic> 16M and the Israeli <jats:italic>B. melitensis<\/jats:italic> field isolates. In addition, we verified their discriminatory power using a set of <jats:italic>B. melitensis<\/jats:italic> strains from various origins and of different MLVA types. Based on our results, we propose MALDI-TOF MS profiling as a rapid, cost-effective alternative to the traditional, time-consuming approach to differentiate certain <jats:italic>B. melitensis<\/jats:italic> isolates on strain level.<\/jats:p>","DOI":"10.3389\/fmicb.2021.712601","type":"journal-article","created":{"date-parts":[[2021,10,22]],"date-time":"2021-10-22T06:18:23Z","timestamp":1634883503000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Protein Biomarker Identification for the Discrimination of Brucella melitensis Field Isolates From the Brucella melitensis Rev.1 Vaccine Strain by MALDI-TOF MS"],"prefix":"10.3389","volume":"12","author":[{"given":"David","family":"Kornspan","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Holger","family":"Brendebach","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dirk","family":"Hofreuter","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shubham","family":"Mathur","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shlomo Eduardo","family":"Blum","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marcelo","family":"Fleker","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Svetlana","family":"Bardenstein","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sascha","family":"Al Dahouk","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2021,10,22]]},"reference":[{"key":"B1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.smallrumres.2016.11.004","article-title":"Duplex PCR for the diagnosis of Brucella melitensis and its differentiation from the REV-1 vaccine strain.","volume":"146","author":"Alvarez","year":"2017","journal-title":"Small Ruminant Res."},{"key":"B2","doi-asserted-by":"publisher","first-page":"232","DOI":"10.3855\/jidc.11949","article-title":"The first report of Brucella melitensis Rev.1 human brucellosis in Bosnia and Herzegovina.","volume":"14","author":"Arapovic","year":"2020","journal-title":"J. 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