Establishment and application of a rapid visual detection method for Listeria monocytogenes based on polymerase spiral reaction (PSR)

Moutong Chen; Tengyi Huang; Min Du; Xiaoxi Bai; Thanapop Soteyome; Lei Yuan; Caiying Bai; Haifeng Lan; Wei Hong; Fang Peng; Xin Fu; Gongyong Peng; Liyan Liu; Birthe V Kjellerup; Zhenbo Xu

doi:10.1080/21655979.2022.2044262

Establishment and application of a rapid visual detection method for Listeria monocytogenes based on polymerase spiral reaction (PSR)

Bioengineered. 2022 Mar;13(3):7860-7867. doi: 10.1080/21655979.2022.2044262.

Authors

Moutong Chen¹, Tengyi Huang², Min Du³, Xiaoxi Bai⁴, Thanapop Soteyome⁵, Lei Yuan⁶, Caiying Bai⁷, Haifeng Lan⁸, Wei Hong³, Fang Peng⁹, Xin Fu³, Gongyong Peng¹⁰, Liyan Liu⁴, Birthe V Kjellerup¹¹, Zhenbo Xu^{2

4

5

11

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Affiliations

¹ Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, Guangdong, China.
² Department of Laboratory Medicine, The Second Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong, China.
³ Gmu-gibh Joint School of Life Sciences, Guangzhou Medical University, Guangzhou, Guangdong, China.
⁴ School of Food Science and Engineering, Guangdong Province Key Laboratory for Green Processing of Natural Products and Product Safety, Engineering Research Center of Starch and Vegetable Protein Processing Ministry of Education, South China University of Technology, Guangzhou, Guangdong, China.
⁵ Home Economics Technology, Rajamangala University of Technology Phra Nakhon, Bangkok, Thailand.
⁶ College of Food Science and Engineering, Yangzhou University, Yangzhou, Jiangsu, China.
⁷ Guangdong Women and Children Hospital, Guangzhou, Guangdong, China.
⁸ Department of Orthopaedic Surgery, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
⁹ Department of Critical Care Medicine, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
¹⁰ State Key Laboratory of Respiratory Diseases, National Clinical Research Center for Respiratory Diseases, National Center for Respiratory Medicine, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
¹¹ Department of Civil and Environmental Engineering, University of Maryland, College Park, MD, USA.
¹² National Institute of Fundamental Studies, Kandy, Sri Lanka.

Abstract

Listeria monocytogenes is a common foodborne pathogen that presents in various food products, posing important threat to public health. The aim of this study was to establish a rapid and sensitive method with visualization to detect L. monocytogenes based on polymerase spiral reaction (PSR). Primers targeting conserved hlyA gene sequence of L. monocytogenes were designed based on bioinformatics analyses on the current available L. monocytogenes genomes. The isothermal amplification PSR can be completed under constant temperature (65ᵒC) within 60 min with high specificity and sensitivity. Twenty-five reference strains were used to evaluate the specificity of the developed reaction. The results showed that the sensitive of the reaction for L. monocytogenes in purified genomic DNA and artificially contaminated food samples were 41 pg/μL and 10³ CFU/mL, respectively. It was 100-fold more sensitive than conventional PCR. In conclusion, this novel PSR method is rapid, cost-efficient, timesaving, and applicable on artificially contaminated food samples, providing broad prospects into the detection of foodborne microbes with the promising on-site inspection.

Keywords: Listeria monocytogenes; food-borne pathogen; hlyA gene; polymerase spiral reaction (PSR); rapid detection.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA Primers / genetics
Food Microbiology
Listeria monocytogenes* / genetics
Nucleic Acid Amplification Techniques / methods
Polymerase Chain Reaction / methods
Sensitivity and Specificity

Substances

DNA Primers

Grants and funding

This work was supported by the State Key Laboratory of Applied Microbiology Southern China (Grant No. SKLAM005-2019), the Open Project of the State Key Laboratory of Respiratory Disease (GHMJLRID-Z-202118), National Natural Science Foundation of China [82170056], the Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program [2017BT01S155], National key Research and Development program of China [2018YFC1311600, 2018YFC1311604, 2016YFC1304100, 2016YFC1304104], Natural Science Foundation of Guangdong (2021A1515011024), Science and Technology Program of Guangzhou (202102080045), The Doctoral Fund of the Third Affiliated Hospital of Guangzhou Medical University (2019B07), Open Project of the State Key Laboratory of Respiratory Disease (SKLRD-Z-202103), Characteristic Innovation Projects of Universities in Guangdong Province (2019KTSCX139), Guangdong Major Project of Basic and Applied Basic Research (2020B0301030005), Guangdong International S&T Cooperation Programme (2021A0505030007).