{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,10]],"date-time":"2026-06-10T07:00:47Z","timestamp":1781074847967,"version":"3.54.1"},"reference-count":31,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T00:00:00Z","timestamp":1777420800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T00:00:00Z","timestamp":1777420800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP24jm0110021"],"award-info":[{"award-number":["JP24jm0110021"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP20wm0125008"],"award-info":[{"award-number":["JP20wm0125008"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP223fa627005"],"award-info":[{"award-number":["JP223fa627005"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbiology and Immunology"],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Glanders is a zoonotic disease caused by\n                    <jats:italic>Burkholderia mallei<\/jats:italic>\n                    (Bm). Bm is phylogenetically similar to\n                    <jats:italic>B. pseudomallei<\/jats:italic>\n                    (Bp), the causative agent of melioidosis, making it difficult to differentiate between them. Recently, the PCR method recommended by the World Organisation for Animal Health (WOAH) failed to detect a Kuwait strain of Bm, and the WOAH has cautioned that ongoing Bm evolution may lead to the emergence of variants that escape standard assays. Therefore, this study aimed to develop a novel real\u2010time PCR method to discriminate between Bm and Bp. Local and web\u2010based BLAST searches identified a 494\u2010bp sequence conserved across Bm but absent from other bacterial genomes, including Bp, except for one\n                    <jats:italic>B. thailandensis<\/jats:italic>\n                    strain. The optimized probe\u2010based real\u2010time PCR targeting this sequence detected the genomes of five Bm isolates but did not amplify DNA from closely related\n                    <jats:italic>Burkholderia<\/jats:italic>\n                    species. In clinical specimens from Mongolia, this assay amplified Bm DNA at an equivalent detection rate to the WOAH\u2010recommended real\u2010time PCR method. These findings provide a novel, highly sensitive diagnostic method for differentiating glanders from melioidosis and suggest its potential use as an alternative method in clinical applications. Further verification using a large number of samples collected from various regions will be necessary in the future.\n                  <\/jats:p>","DOI":"10.1111\/1348-0421.70061","type":"journal-article","created":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T05:20:07Z","timestamp":1777526407000},"page":"305-314","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a Novel Real\u2010Time PCR Assay Differentiating\n                    <i>Burkholderia mallei<\/i>\n                    From\n                    <i>Burkholderia pseudomallei<\/i>"],"prefix":"10.1111","volume":"70","author":[{"given":"Ai","family":"Koshikawa","sequence":"first","affiliation":[{"name":"Laboratory of Comparative Pathology, Faculty of Veterinary Medicine Hokkaido University Sapporo Hokkaido Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshiki","family":"Ichikawa","sequence":"additional","affiliation":[{"name":"Department of Veterinary Forensics, Faculty of Veterinary Medicine Hokkaido University Sapporo Hokkaido Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mitsuru","family":"Nakase","sequence":"additional","affiliation":[{"name":"Laboratory of Comparative Pathology, Faculty of Veterinary Medicine Hokkaido University Sapporo Hokkaido Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Apichai","family":"Tuanyok","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases and Immunology, College of Veterinary Medicine University of Florida Gainesville Florida USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vannarat","family":"Saechan","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Science Prince of Songkla University Songkhla Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keisuke","family":"Aoshima","sequence":"additional","affiliation":[{"name":"Laboratory of Comparative Pathology, Faculty of Veterinary Medicine Hokkaido University Sapporo Hokkaido Japan"},{"name":"Cancer Research Unit, One Health Research Center Hokkaido University Sapporo Hokkaido Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hideaki","family":"Higashi","sequence":"additional","affiliation":[{"name":"Division of Infection and Immunity, International Institute for Zoonosis Control Hokkaido University Sapporo Hokkaido Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chie","family":"Nakajima","sequence":"additional","affiliation":[{"name":"Division of Bioresources, International Institute for Zoonosis Control Hokkaido University Sapporo Hokkaido Japan"},{"name":"Institute for Vaccine Research and Development Hokkaido University Sapporo Hokkaido Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasuhiko","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Division of Bioresources, International Institute for Zoonosis Control Hokkaido University Sapporo Hokkaido Japan"},{"name":"Institute for Vaccine Research and Development Hokkaido University Sapporo Hokkaido Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanaabaatar","family":"Batbaatar","sequence":"additional","affiliation":[{"name":"Laboratory of Infectious Disease and Immunology, Institute of Veterinary Medicine Mongolian University of Life Sciences Zaisan Ulaanbaatar Mongolia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7286-8365","authenticated-orcid":false,"given":"Takashi","family":"Kimura","sequence":"additional","affiliation":[{"name":"Laboratory of Comparative Pathology, Faculty of Veterinary Medicine Hokkaido University Sapporo Hokkaido Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,29]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1348-0421.1992.tb02129.x"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1865-1682.2012.01342.x"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/26089"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.3201\/eid0802.010164"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1186\/1746-6148-8-214"},{"key":"e_1_2_12_7_1","volume-title":"Veterinary Medicine: A textbook of the Diseases of Cattle, Horses, Sheep, Pigs, and Goats","author":"Peter C. 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