ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Food Microbiology
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1596797
This article is part of the Research TopicCritical- and High-Priority Pathogens in the Food ChainView all 5 articles
Rapid and simple detection of Listeria monocytogenes using real closed dumbbell mediated isothermal amplification (CDA)
Provisionally accepted
- 1Department of Hematology, Ningbo First Hospital, Ningbo, Zhejiang Province, China
- 2Department of Experimental Medical Science, Ningbo No. 2 Hospital, Ningbo, China
- 3Zhenhai People's Hospital, Ningbo, Zhejiang Province, China
- 4Ningbo First Hospital, Ningbo, Zhejiang Province, China
Select one of your emails
You have multiple emails registered with Frontiers:
Notify me on publication
Please enter your email address:
If you already have an account, please login
You don't have a Frontiers account ? You can register here
Introduction: Listeria monocytogenes (L. monocytogenes) is a well-known widespread food-borne pathogen which posed a threaten to public health. Suitable detection methods are needed to effectively control and prevent pathogenic L. monocytogenes infections.Methods: This study was to develop a novel closed dumbbell mediated isothermal amplification (CDA) based assay to achieve rapid and simple detection of L. monocytogenes. The newly-developed CDA technology capable of amplifying DNA target with high sensitivity and specificity. The conserved hly gene of L. monocytogenes was taken as target for establishment of CDA method. All primers were selected and evaluated by real-time fluorescence monitoring and endpoint visual judgement indicated by hydroxy naphthol blue (HNB). Results: The specificity and sensitivity of this CDA based diagnostic system was determined after the evaluation of 560 batches of DNA samples detection. The detection limit of the L. monocytogenes O-CDA assay was 1 copy/μl using artificial samples. The results of real-time fluorescence-based O-CDA coupled with melting curves analysis showed the method would achieve rapid and accurate diagnosis of L. monocytogenes, can be employed as an alternative to qPCR in diagnostic practice. Moreover, the L. monocytogenes O-CDA method monitored by real-time fluorescence and endpoint hydroxy naphthol blue (HNB) based colorimetric displayed same sensitivity, specificity and accuracy, would be helpful to realize onsite pathogen surveillance. Discussion: The real-time fluorescence plots and following melting curve analysis-based L. monocytogenes O-CDA were suitable for laboratory-based. Considering the portable manipulation of HNB based colorimetric detection system, our results shed light on its potential application for on-site L. monocytogenes surveillance. The developed CDA based methods are rapid, simple, reliable, and sensitive in several samples, showed potential to manage the task of L. monocytogenes monitoring easier.
Keywords: Listeria monocytogenes, Point of care diagnostic, Closed dumbbell mediated isothermal amplification (CDA), Real-time fluorescence CDA, Visual CDA
Received: 20 Mar 2025; Accepted: 09 Jun 2025.
Copyright: © 2025 Zhang, Wu, Zhong, Xuhan, Fei, Ouyang and Mao. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Rui Mao, Department of Experimental Medical Science, Ningbo No. 2 Hospital, Ningbo, China
Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.