{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T15:55:13Z","timestamp":1789487713852,"version":"build-2803163510"},"reference-count":14,"publisher":"Oxford University Press (OUP)","issue":"9","license":[{"start":{"date-parts":[[2026,8,22]],"date-time":"2026-08-22T00:00:00Z","timestamp":1787356800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The xerophilic fungal genus Wallemia is an important cause of fungal contamination in bread manufacturing environments, but its detection by conventional culture-based methods is often difficult because of slow growth and competitive overgrowth by other fungi. Here, we report the development of a genus-specific TaqMan real-time polymerase chain reaction assay targeting the internal transcribed spacer (ITS) region for rapid detection of Wallemia. Primer and probe sequences were designed based on comparative ITS analysis of all nine recognized Wallemia species and non-target fungi commonly encountered in bakery environments. The probe-binding region was fully conserved among all Wallemia ITS reference sequences evaluated. Analytical specificity was confirmed using synthetic DNA fragments representing predicted amplicon groups, with amplification observed exclusively in Wallemia. The assay demonstrated excellent linearity over 10\u00b2\u201310\u2076 copies per reaction (R\u00b2 = 0.9988), with an amplification efficiency of 102.7%. Analytical sensitivity evaluation yielded an estimated LOD95 of 9.46 copies per reaction. Application to environmental swab samples from an operational bakery production line enabled culture-independent detection of Wallemia, including in samples in which culture-based identification was obscured by fungal overgrowth. This assay provides a rapid and specific molecular tool for culture-independent monitoring of Wallemia contamination and may support improved hygienic control in bakery manufacturing environments.<\/jats:p>","DOI":"10.1093\/lambio\/ovag072","type":"journal-article","created":{"date-parts":[[2026,8,22]],"date-time":"2026-08-22T11:39:38Z","timestamp":1787398778000},"source":"Crossref","is-referenced-by-count":0,"title":["A genus-specific TaqMan real-time polymerase chain reaction assay for the rapid detection of\n                    <i>Wallemia<\/i>\n                    in bakery production environments"],"prefix":"10.1093","volume":"79","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2504-2894","authenticated-orcid":false,"given":"Akira","family":"Aoki","sequence":"first","affiliation":[{"name":"Faculty of Pharmacy, Meijo University , 150 Yagotoyama, Tempaku-ku, Nagoya 468-8503 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Megumi","family":"Watanabe","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy, Meijo University , 150 Yagotoyama, Tempaku-ku, Nagoya 468-8503 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Asuka","family":"Maruo","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy, Meijo University , 150 Yagotoyama, Tempaku-ku, Nagoya 468-8503 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Machiko","family":"Kuchiji","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy, Meijo University , 150 Yagotoyama, Tempaku-ku, Nagoya 468-8503 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshinori","family":"Okamoto","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy, Meijo University , 150 Yagotoyama, Tempaku-ku, Nagoya 468-8503 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,8,22]]},"reference":[{"key":"2026091510313651100_bib1","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments","volume":"55","author":"Bustin","year":"2009","journal-title":"Clin Chem"},{"key":"2026091510313651100_bib2","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1016\/S0167-7012(02)00037-4","article-title":"Evaluation of rapid DNA extraction methods for the quantitative detection of fungi using real-time PCR analysis","volume":"50","author":"Haugland","year":"2002","journal-title":"J Microbiol Meth"},{"key":"2026091510313651100_bib3","doi-asserted-by":"publisher","first-page":"986","DOI":"10.1101\/gr.6.10.986","article-title":"Real time quantitative PCR","volume":"6","author":"Heid","year":"1996","journal-title":"Genome Res"},{"key":"2026091510313651100_bib4","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1007\/s13225-015-0333-x","article-title":"Halophily reloaded: new insights into the extremophilic life-style of Wallemia with the description of Wallemia hederae sp. nov","volume":"76","author":"Jan\u010di\u010d","year":"2016","journal-title":"Fung Divers"},{"key":"2026091510313651100_bib5","doi-asserted-by":"publisher","first-page":"772","DOI":"10.1093\/molbev\/mst010","article-title":"MAFFT multiple sequence alignment software version 7: improvements in performance and usability","volume":"30","author":"Katoh","year":"2013","journal-title":"Molecul Biol Evol"},{"key":"2026091510313651100_bib6","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1016\/0964-8305(93)90038-4","article-title":"Mould spoilage of bread: the problem and some solutions","volume":"32","author":"Legan","year":"1993","journal-title":"Int Biodeterior Biodegrad"},{"key":"2026091510313651100_bib7","doi-asserted-by":"publisher","first-page":"190","DOI":"10.1111\/j.1198-743X.2004.00722.x","article-title":"Real-time PCR in the microbiology laboratory","volume":"10","author":"Mackay","year":"2004","journal-title":"Clin Microbiol Infect"},{"key":"2026091510313651100_bib8","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/j.ijfoodmicro.2007.07.034","article-title":"Post-harvest control strategies: minimizing mycotoxins in the food chain","volume":"119","author":"Magan","year":"2007","journal-title":"Int J Food Microbiol"},{"key":"2026091510313651100_bib9","doi-asserted-by":"publisher","first-page":"25","DOI":"10.3358\/shokueishi.55.25","article-title":"Development of direct real-time PCR system applicable to a wide range of foods and agricultural products","volume":"55","author":"Mano","year":"2014","journal-title":"J Food Hyg Soc Jpn"},{"key":"2026091510313651100_bib10","doi-asserted-by":"publisher","first-page":"1853","DOI":"10.1128\/am.16.12.1853-1858.1968","article-title":"Water relations of xerophilic fungi isolated from prunes","volume":"16","author":"Pitt","year":"1968","journal-title":"Appl Microbiol"},{"key":"2026091510313651100_bib11","doi-asserted-by":"publisher","first-page":"848","DOI":"10.1016\/j.fm.2011.02.008","article-title":"Recent advances in quantitative PCR (qPCR) applications in food microbiology","volume":"28","author":"Postollec","year":"2011","journal-title":"Food Microbiol"},{"key":"2026091510313651100_bib12","doi-asserted-by":"publisher","first-page":"6241","DOI":"10.1073\/pnas.1117018109","article-title":"Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi","volume":"109","author":"Schoch","year":"2012","journal-title":"Proc Natl Acad Sci USA"},{"key":"2026091510313651100_bib13","doi-asserted-by":"publisher","first-page":"E46","DOI":"10.3390\/microorganisms6020046","article-title":"The genus Wallemia-from contamination of food to health threat","volume":"6","author":"Zajc","year":"2018","journal-title":"Microorganisms"},{"key":"2026091510313651100_bib14","doi-asserted-by":"publisher","first-page":"7295","DOI":"10.1128\/AEM.70.12.7295-7302.2004","article-title":"Detection and quantification of Wallemia sebi in aerosols by real-time PCR, conventional PCR, and cultivation","volume":"70","author":"Zeng","year":"2004","journal-title":"Appl Environ Microb"}],"container-title":["Letters in Applied Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/lambio\/advance-article-pdf\/doi\/10.1093\/lambio\/ovag072\/70763838\/ovag072.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/lambio\/article-pdf\/79\/9\/ovag072\/70763838\/ovag072.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/lambio\/article-pdf\/79\/9\/ovag072\/70763838\/ovag072.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T14:31:43Z","timestamp":1789482703000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/lambio\/article\/doi\/10.1093\/lambio\/ovag072\/8768683"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,22]]},"references-count":14,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2026,9,4]]}},"URL":"https:\/\/doi.org\/10.1093\/lambio\/ovag072","relation":{},"ISSN":["1472-765X"],"issn-type":[{"value":"1472-765X","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2026,9]]},"published":{"date-parts":[[2026,8,22]]},"article-number":"ovag072"}}