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Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Pseudomonas fluorescens in Raw Milk
Raw milk is susceptible to microbial contamination during transportation and storage. Pseudomonas fluorescens producing heat-resistant enzymes have become the most common and harmful psychrophilic microorganisms in the cold chain logistics of raw milk. To rapidly detect P. fluorescens in raw milk, t...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8770903/ https://www.ncbi.nlm.nih.gov/pubmed/35069513 http://dx.doi.org/10.3389/fmicb.2021.810511 |
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author | Bu, Yushan Qiao, Wenjun Zhai, Zhengyuan Liu, Tongjie Gong, Pimin Zhang, Lanwei Hao, Yanling Yi, Huaxi |
author_facet | Bu, Yushan Qiao, Wenjun Zhai, Zhengyuan Liu, Tongjie Gong, Pimin Zhang, Lanwei Hao, Yanling Yi, Huaxi |
author_sort | Bu, Yushan |
collection | PubMed |
description | Raw milk is susceptible to microbial contamination during transportation and storage. Pseudomonas fluorescens producing heat-resistant enzymes have become the most common and harmful psychrophilic microorganisms in the cold chain logistics of raw milk. To rapidly detect P. fluorescens in raw milk, the protease gene aprX was selected as a detection target to construct a set of primers with strong specificity, and a loop-mediated isothermal amplification (LAMP) assay was established. The detection thresholds of the LAMP assay for pure cultured P. fluorescens and pasteurized milk were 2.57 × 10(2) and 3 × 10(2) CFU/mL, respectively. It had the advantages over conventional method of low detection threshold, strong specificity, rapid detection, and simple operation. This LAMP assay can be used for online monitoring and on-site detection of P. fluorescens in raw milk to guarantee the quality and safety of dairy products. |
format | Online Article Text |
id | pubmed-8770903 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-87709032022-01-21 Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Pseudomonas fluorescens in Raw Milk Bu, Yushan Qiao, Wenjun Zhai, Zhengyuan Liu, Tongjie Gong, Pimin Zhang, Lanwei Hao, Yanling Yi, Huaxi Front Microbiol Microbiology Raw milk is susceptible to microbial contamination during transportation and storage. Pseudomonas fluorescens producing heat-resistant enzymes have become the most common and harmful psychrophilic microorganisms in the cold chain logistics of raw milk. To rapidly detect P. fluorescens in raw milk, the protease gene aprX was selected as a detection target to construct a set of primers with strong specificity, and a loop-mediated isothermal amplification (LAMP) assay was established. The detection thresholds of the LAMP assay for pure cultured P. fluorescens and pasteurized milk were 2.57 × 10(2) and 3 × 10(2) CFU/mL, respectively. It had the advantages over conventional method of low detection threshold, strong specificity, rapid detection, and simple operation. This LAMP assay can be used for online monitoring and on-site detection of P. fluorescens in raw milk to guarantee the quality and safety of dairy products. Frontiers Media S.A. 2022-01-06 /pmc/articles/PMC8770903/ /pubmed/35069513 http://dx.doi.org/10.3389/fmicb.2021.810511 Text en Copyright © 2022 Bu, Qiao, Zhai, Liu, Gong, Zhang, Hao and Yi. https://creativecommons.org/licenses/by/4.0/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) and the copyright owner(s) 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. |
spellingShingle | Microbiology Bu, Yushan Qiao, Wenjun Zhai, Zhengyuan Liu, Tongjie Gong, Pimin Zhang, Lanwei Hao, Yanling Yi, Huaxi Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Pseudomonas fluorescens in Raw Milk |
title | Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Pseudomonas fluorescens in Raw Milk |
title_full | Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Pseudomonas fluorescens in Raw Milk |
title_fullStr | Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Pseudomonas fluorescens in Raw Milk |
title_full_unstemmed | Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Pseudomonas fluorescens in Raw Milk |
title_short | Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Pseudomonas fluorescens in Raw Milk |
title_sort | establishment and evaluation of a loop-mediated isothermal amplification assay for rapid detection of pseudomonas fluorescens in raw milk |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8770903/ https://www.ncbi.nlm.nih.gov/pubmed/35069513 http://dx.doi.org/10.3389/fmicb.2021.810511 |
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