AUTHOR=Bu Yushan , Qiao Wenjun , Zhai Zhengyuan , Liu Tongjie , Gong Pimin , Zhang Lanwei , Hao Yanling , Yi Huaxi 

TITLE=Establishment and Evaluation of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Pseudomonas fluorescens in Raw Milk

JOURNAL=Frontiers in Microbiology

VOLUME=12

YEAR=2022

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.810511

DOI=10.3389/fmicb.2021.810511

ISSN=1664-302X

ABSTRACT=<p>Raw milk is susceptible to microbial contamination during transportation and storage. <italic>Pseudomonas fluorescens</italic> producing heat-resistant enzymes have become the most common and harmful psychrophilic microorganisms in the cold chain logistics of raw milk. To rapidly detect <italic>P. fluorescens</italic> in raw milk, the protease gene <italic>apr</italic>X 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 <italic>P. fluorescens</italic> and pasteurized milk were 2.57 × 10<sup>2</sup> and 3 × 10<sup>2</sup> 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 <italic>P. fluorescens</italic> in raw milk to guarantee the quality and safety of dairy products.</p>