{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T14:21:55Z","timestamp":1789222915204,"version":"build-2803163510"},"reference-count":44,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,8,19]],"date-time":"2025-08-19T00:00:00Z","timestamp":1755561600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Listeria monocytogenes<\/jats:italic>\n                      (\n                      <jats:italic>L. monocytogenes<\/jats:italic>\n                      ) is a well-known widespread food-borne pathogen that poses a threat to public health. Suitable detection methods are needed to effectively control and prevent pathogenic\n                      <jats:italic>L. monocytogenes<\/jats:italic>\n                      infections.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      This study aimed to develop a novel closed dumbbell-mediated isothermal amplification (CDA)-based assay to achieve rapid and simple detection of\n                      <jats:italic>L. monocytogenes.<\/jats:italic>\n                      The newly developed CDA technology is capable of amplifying DNA targets with high sensitivity and specificity. The conserved\n                      <jats:italic>hly<\/jats:italic>\n                      gene of\n                      <jats:italic>L. monocytogenes<\/jats:italic>\n                      was taken as a target for the establishment of the CDA method. All primers were selected and evaluated by real-time fluorescence monitoring, and endpoint visual judgement was indicated by hydroxy naphthol blue (HNB).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The specificity and sensitivity of this CDA-based diagnostic system were determined after the evaluation of 560 batches of DNA samples. The detection limit of the\n                      <jats:italic>L. monocytogenes<\/jats:italic>\n                      O-CDA assay was 1 copy\/\u03bcl using artificial samples. The results of real-time fluorescence-based O-CDA coupled with melting curves analysis showed that the method would achieve rapid and accurate diagnosis of\n                      <jats:italic>L. monocytogenes<\/jats:italic>\n                      , and can be employed as an alternative to qPCR in diagnostic practice. Moreover, the\n                      <jats:italic>L. monocytogenes<\/jats:italic>\n                      O-CDA method monitored by real-time fluorescence and endpoint hydroxy naphthol blue (HNB) based colorimetric assay displayed the same sensitivity, specificity, and accuracy, which would be helpful to realize onsite pathogen surveillance.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      The real-time fluorescence plots and following melting curve analysis-based\n                      <jats:italic>L. monocytogenes<\/jats:italic>\n                      O-CDA were suitable for laboratory-based diagnosis. Considering the portable manipulation of the HNB-based colorimetric detection system, our results shed light on its potential application for on-site\n                      <jats:italic>L. monocytogenes<\/jats:italic>\n                      surveillance. The developed CDA-based methods are rapid, simple, reliable, and sensitive in several samples, showing potential to manage the task of\n                      <jats:italic>L. monocytogenes<\/jats:italic>\n                      monitoring more easily.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2025.1596797","type":"journal-article","created":{"date-parts":[[2025,8,19]],"date-time":"2025-08-19T05:20:29Z","timestamp":1755580829000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Rapid and simple detection of Listeria monocytogenes using real closed dumbbell-mediated isothermal amplification"],"prefix":"10.3389","volume":"16","author":[{"given":"Yanli","family":"Zhang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyao","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yeling","family":"Zhong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuhan","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fei","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guifang","family":"Ouyang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rui","family":"Mao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,8,19]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"675","DOI":"10.1089\/fpd.2020.2923","article-title":"Detection and virulence characterization of Listeria monocytogenes strains in ready-to-eat products","volume":"18","author":"Amagliani","year":"2021","journal-title":"Foodborne Pathog. 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