{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T11:02:54Z","timestamp":1787828574087,"version":"build-2784847793"},"reference-count":50,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2025,4,1]],"date-time":"2025-04-01T00:00:00Z","timestamp":1743465600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,4,1]],"date-time":"2025-04-01T00:00:00Z","timestamp":1743465600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2025,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              <jats:italic>Listeria monocytogenes<\/jats:italic>\n              , a rod-shaped Gram-positive bacterium widely distributed in nature, can contaminate foods and represents a foodborne pathogen of public health significance causing a high mortality rate of 20%\u201330%. Rapid and reliable identification of foods and food-processing environments contaminated with\n              <jats:italic>L. monocytogenes<\/jats:italic>\n              is a crucial step in implementing effective intervention strategies to ensure food safety and limit the transmission of bacteria to humans. This study designed and refined a practical workflow to streamline and accelerate the detection of a low level of\n              <jats:italic>L. monocytogenes<\/jats:italic>\n              present in ground beef. The workflow coupled an abbreviated 5 h culture enrichment in PALCAM liquid medium with physical separation (filtration and centrifugation) to preprocess enrichment samples. Specific capture was achieved using magnetic separation with a bacteriophage endolysin-derived cell wall-binding domain in a Hyglos\n              <jats:italic>Listeria<\/jats:italic>\n              capture kit. Molecular detection was performed using a MicroSEQ\n              <jats:italic>L. monocytogenes<\/jats:italic>\n              RTi-PCR detection kit combined with a nested PCR strategy. Preprocessing of enrichment culture samples using a multi-stage filtration system constructed for the study or commercially available BagFilter Pull-up filter bags, in conjunction with centrifugation, enabled the recovery of ~30 colony-forming units (CFUs) from the enrichment culture of a 25 g ground beef sample artificially contaminated with 1 CFU of\n              <jats:italic>L. monocytogenes<\/jats:italic>\n              . Integration of magnetic separation into the workflow for capturing\n              <jats:italic>L. monocytogenes<\/jats:italic>\n              cells specifically from preprocessed samples and further cleaning up the samples yielded bacterial counts similar to those obtained by direct plating of preprocessed samples. The RTi-PCR-based molecular detection method integrated into the workflow was capable of detecting pure cultures of\n              <jats:italic>L. monocytogenes<\/jats:italic>\n              as low as 12.5 CFUs. Evaluation of the workflow using artificially ground beef demonstrated the consistent detection of\n              <jats:italic>L. monocytogenes<\/jats:italic>\n              within an 8 h workday in a 25 g sample unit containing the cell count as low as 2 CFU following a 5 h culture enrichment.\n            <\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>\n              Consuming foods contaminated with the bacterial pathogen\n              <jats:italic>Listeria monocytogenes<\/jats:italic>\n              can lead to the development of human listeriosis, a severe and life-threatening foodborne illness. Timely detection of\n              <jats:italic>L. monocytogenes<\/jats:italic>\n              present at a low level in foods and food processing environments is a necessary measure to prevent the spread of the\n              <jats:italic>Listeria<\/jats:italic>\n              -associated illness. This study designed and evaluated a multi-step workflow for testing\n              <jats:italic>L. monocytogenes<\/jats:italic>\n              in artificially contaminated food samples. The workflow was composed of a short 5 h culture enrichment, filtration-based sample preprocessing, magnetic separation, a single-tube nested RTi-PCR, and culture plating. It allowed\n              <jats:italic>L. monocytogenes<\/jats:italic>\n              to be detected within 8 h from a 25 g ground beef sample containing the target cells as low as 2 colony-forming units, significantly improving and streamlining the detection methods for this important foodborne pathogen.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/spectrum.01577-24","type":"journal-article","created":{"date-parts":[[2025,2,25]],"date-time":"2025-02-25T14:00:42Z","timestamp":1740492042000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":6,"title":["A streamlined procedure for advancing the detection and isolation of\n            <i>Listeria monocytogenes<\/i>\n            from artificially contaminated ground beef in a single working day"],"prefix":"10.1128","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7351-6046","authenticated-orcid":true,"given":"Min","family":"Lin","sequence":"first","affiliation":[{"name":"Canadian Food Inspection Agency, Ottawa Laboratory Fallowfield","place":["Ottawa, Canada"]},{"name":"Department of Biochemistry, Microbiology and Immunology, University of Ottawa","place":["Ottawa, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanhong","family":"Dan","sequence":"additional","affiliation":[{"name":"Canadian Food Inspection Agency, Ottawa Laboratory Fallowfield","place":["Ottawa, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiewen","family":"Guan","sequence":"additional","affiliation":[{"name":"Canadian Food Inspection Agency, Ottawa Laboratory Fallowfield","place":["Ottawa, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2007.05.011"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-016-7552-2"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1601.091155"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0505.990502"},{"key":"e_1_3_2_6_2","unstructured":"World Health Organization. 2018. 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