{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T12:14:55Z","timestamp":1772540095951,"version":"3.50.1"},"reference-count":33,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,8,2]],"date-time":"2025-08-02T00:00:00Z","timestamp":1754092800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"TED MAPA\/EMBRAPA, Technical-Scientific Cooperation Agreement ABQM\/EMBRAPA","award":["10.23.10.011.00.00"],"award-info":[{"award-number":["10.23.10.011.00.00"]}]},{"name":"TED MAPA\/EMBRAPA, Technical-Scientific Cooperation Agreement ABQM\/EMBRAPA","award":["315857\/2021-8"],"award-info":[{"award-number":["315857\/2021-8"]}]},{"name":"CNPq","award":["10.23.10.011.00.00"],"award-info":[{"award-number":["10.23.10.011.00.00"]}]},{"name":"CNPq","award":["315857\/2021-8"],"award-info":[{"award-number":["315857\/2021-8"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Glanders is a highly contagious and often fatal zoonotic disease of equids caused by Burkholderia mallei, a pathogen of significant concern due to its potential for bioterrorism. In Brazil, glanders remains endemic, particularly among working equids in the Northeast region. Diagnostic confirmation typically involves serology, culture, and polymerase chain reaction (PCR), although false-negative PCR results have been increasingly reported. This study aimed to evaluate the diagnostic performance and analytical sensitivity of four B. mallei-specific PCR primer sets using samples from 30 seropositive equids. Microbiological cultures were obtained from various organs and swabs, followed by PCR targeting four genomic regions: fliP-IS407A(a), fliP-IS407A(b), Burk457, and Bm17. All animals were confirmed positive for B. mallei via culture, but PCR detection rates varied significantly across primer sets. The fliP-IS407A(b) primer set showed the highest sensitivity, detecting 86% of samples, while the WOAH-recommended fliP-IS407A(a) set had the lowest performance (13.4%). Analytical sensitivity assays confirmed that fliP-IS407A(b) and Bm17 primers detected DNA concentrations as low as 0.007 ng, outperforming the others. These findings suggest that certain widely used primer sets may lack sufficient sensitivity for reliable detection of B. mallei, especially in chronically infected animals with low bacterial loads. The study underscores the need for ongoing validation of molecular diagnostics to improve the detection and control of glanders in endemic regions.<\/jats:p>","DOI":"10.3390\/pathogens14080766","type":"journal-article","created":{"date-parts":[[2025,8,6]],"date-time":"2025-08-06T07:45:11Z","timestamp":1754466311000},"page":"766","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Detection of Burkholderia mallei in Microbiological Culture: A Comparative Analysis of PCR Primer Sets"],"prefix":"10.3390","volume":"14","author":[{"given":"J\u00e9ssica Cristine K.","family":"Moriya","sequence":"first","affiliation":[{"name":"CNPq Scholarship\/Embrapa Beef Cattle, Campo Grande 79106-550, MS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paula Adas P.","family":"Suniga","sequence":"additional","affiliation":[{"name":"Embrapa Beef Cattle\/Ministry of Agriculture, Livestock and Food Supply Scholarship, Campo Grande 79106-550, MS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana Clara L.","family":"Ara\u00fajo","sequence":"additional","affiliation":[{"name":"CNPq\/PIBIC Program\/Embrapa Beef Cattle, Campo Grande 79106-550, MS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Goretti","family":"Santos","sequence":"additional","affiliation":[{"name":"Embrapa Beef Cattle, Campo Grande 79106-550, MS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juliana S. G.","family":"Rieger","sequence":"additional","affiliation":[{"name":"Embrapa Beef Cattle\/Ministry of Agriculture, Livestock and Food Supply Scholarship, Campo Grande 79106-550, MS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cynthia","family":"Mantovani","sequence":"additional","affiliation":[{"name":"Embrapa Beef Cattle\/Ministry of Agriculture, Livestock and Food Supply Scholarship, Campo Grande 79106-550, MS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0943-5356","authenticated-orcid":false,"given":"Rodrigo","family":"Jardim","sequence":"additional","affiliation":[{"name":"Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro 21040-900, RJ, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M\u00e1rcio Roberto","family":"Silva","sequence":"additional","affiliation":[{"name":"Embrapa Dairy Cattle, Juiz de Fora 36038-330, MG, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fl\u00e1bio R.","family":"Ara\u00fajo","sequence":"additional","affiliation":[{"name":"Embrapa Beef Cattle, Campo Grande 79106-550, MS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lenita R.","family":"Santos","sequence":"additional","affiliation":[{"name":"Embrapa Beef Cattle, Campo Grande 79106-550, MS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"654","DOI":"10.1111\/evj.12599","article-title":"Glanders and the risk for its introduction through the international movement of horses","volume":"48","author":"Kettle","year":"2016","journal-title":"Equine Vet. 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