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Detection of Listeria monocytogenes in Food Using the Proofman-LMTIA Assay

Microbial factors, including bacteria, viruses, and other pathogens, are significant contributors to foodborne illnesses, posing serious food safety risks due to their potential for rapid growth and contamination. Listeria monocytogenes is one of the most common types of foodborne bacteria that can...

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Autores principales: Song, Chunmei, Wang, Borui, Wang, Yongzhen, Liu, Jinxin, Wang, Deguo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10385859/
https://www.ncbi.nlm.nih.gov/pubmed/37513329
http://dx.doi.org/10.3390/molecules28145457
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author Song, Chunmei
Wang, Borui
Wang, Yongzhen
Liu, Jinxin
Wang, Deguo
author_facet Song, Chunmei
Wang, Borui
Wang, Yongzhen
Liu, Jinxin
Wang, Deguo
author_sort Song, Chunmei
collection PubMed
description Microbial factors, including bacteria, viruses, and other pathogens, are significant contributors to foodborne illnesses, posing serious food safety risks due to their potential for rapid growth and contamination. Listeria monocytogenes is one of the most common types of foodborne bacteria that can cause serious foodborne diseases or even fatalities. In this study, a novel nucleic acid amplification method called Proofman-LMTIA was employed to detect Listeria monocytogenes contamination in food. This method combines proofreading enzyme-mediated probe cleavage with ladder-shape melting temperature isothermal amplification. A positive recombinant plasmid was used as a control to ensure the accuracy of the detection results, and primers and Proofman probes were specifically designed for the LMTIA. Genomic DNA was extracted, the reaction temperature was optimized, and the primers’ specificity was verified using foodborne pathogens like Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella. The sensitivity was assessed by testing serial dilutions of genomic DNA, and the method’s applicability was confirmed by detecting artificially contaminated fresh pork. The established LMTIA method exhibited both high specificity and sensitivity. At the optimal reaction temperature of 63 °C, the primers specifically identified Listeria monocytogenes contamination in pork at a concentration of 8.0 ± 0.7 colony-forming units (CFUs) per 25 g. Furthermore, the Proofman-LMTIA method was applied to test Listeria monocytogenes DNA in 30 food samples purchased from a Chinese retail market, and reassuringly, all results indicated no contamination. Proofman-LMTIA can serve as a reliable and rapid method for detecting Listeria monocytogenes in food, contributing to public health by safeguarding consumers from foodborne illnesses, and strengthening food safety regulations.
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spelling pubmed-103858592023-07-30 Detection of Listeria monocytogenes in Food Using the Proofman-LMTIA Assay Song, Chunmei Wang, Borui Wang, Yongzhen Liu, Jinxin Wang, Deguo Molecules Article Microbial factors, including bacteria, viruses, and other pathogens, are significant contributors to foodborne illnesses, posing serious food safety risks due to their potential for rapid growth and contamination. Listeria monocytogenes is one of the most common types of foodborne bacteria that can cause serious foodborne diseases or even fatalities. In this study, a novel nucleic acid amplification method called Proofman-LMTIA was employed to detect Listeria monocytogenes contamination in food. This method combines proofreading enzyme-mediated probe cleavage with ladder-shape melting temperature isothermal amplification. A positive recombinant plasmid was used as a control to ensure the accuracy of the detection results, and primers and Proofman probes were specifically designed for the LMTIA. Genomic DNA was extracted, the reaction temperature was optimized, and the primers’ specificity was verified using foodborne pathogens like Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella. The sensitivity was assessed by testing serial dilutions of genomic DNA, and the method’s applicability was confirmed by detecting artificially contaminated fresh pork. The established LMTIA method exhibited both high specificity and sensitivity. At the optimal reaction temperature of 63 °C, the primers specifically identified Listeria monocytogenes contamination in pork at a concentration of 8.0 ± 0.7 colony-forming units (CFUs) per 25 g. Furthermore, the Proofman-LMTIA method was applied to test Listeria monocytogenes DNA in 30 food samples purchased from a Chinese retail market, and reassuringly, all results indicated no contamination. Proofman-LMTIA can serve as a reliable and rapid method for detecting Listeria monocytogenes in food, contributing to public health by safeguarding consumers from foodborne illnesses, and strengthening food safety regulations. MDPI 2023-07-17 /pmc/articles/PMC10385859/ /pubmed/37513329 http://dx.doi.org/10.3390/molecules28145457 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Song, Chunmei
Wang, Borui
Wang, Yongzhen
Liu, Jinxin
Wang, Deguo
Detection of Listeria monocytogenes in Food Using the Proofman-LMTIA Assay
title Detection of Listeria monocytogenes in Food Using the Proofman-LMTIA Assay
title_full Detection of Listeria monocytogenes in Food Using the Proofman-LMTIA Assay
title_fullStr Detection of Listeria monocytogenes in Food Using the Proofman-LMTIA Assay
title_full_unstemmed Detection of Listeria monocytogenes in Food Using the Proofman-LMTIA Assay
title_short Detection of Listeria monocytogenes in Food Using the Proofman-LMTIA Assay
title_sort detection of listeria monocytogenes in food using the proofman-lmtia assay
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10385859/
https://www.ncbi.nlm.nih.gov/pubmed/37513329
http://dx.doi.org/10.3390/molecules28145457
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