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Development of a Rapid Screening Test for Listeria monocytogenes in Raw Chicken Meat Using Loop-Mediated Isothermal Amplification (LAMP) and Lateral Flow Dipstick (LFD)

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Abstract

A loop-mediated isothermal amplification and lateral flow dipstick (LAMP-LFD) protocol was established for the rapid detection of the foodborne pathogen Listeria monocytogenes based on detection of the phosphatidylcholine-phospholipase C gene (plcB). The LAMP-LFD method achieved results within 90 min and consisted of a one-step DNA extraction followed by LAMP and LFD. The detection limits of the assay using L. monocytogenes purified genomic DNA or cells from a pure culture were 800 fg and 2.82 CFU mL−1, respectively. The specificity test revealed that the method exhibited no cross-reactivity with other Listeria species (Listeria innocua DMST 9011, Listeria ivanovii DMST 9012, Listeria welshimeri DMST 20559) or non-Listeria spp. (Salmonella ssp., Shigella spp., Campylobacter spp., Escherichia coli ATCC 25922, Bacillus cereus, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Micrococcus luteus, Citrobacter diversus, Serratia marcescens, Enterobacter aerogenes, and Klebsiella oxytoca). Compared to standard culture analysis, the specificity, sensitivity, and accuracy of the LAMP-LFD method in tests of 200 raw chicken meat samples were 100, 90.20, and 97.50%, respectively. Thus, LAMP-LFD is a promising point-of-care diagnosis tool for decision-making for controlling of L. monocytogenes-contaminated food products, decreasing mortality rates, and improving the quality of life.

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Acknowledgements

The research funding was supported by Thailand Research Fund through the Royal Golden Jubilee Ph.D. Program (Grant No. PHD/0040/2554), The Graduate school of Srinakharinwirot University, and Agricultural Research Development Agency (Public organization) (No. CRP5705022280), Thailand. The authors are grateful to all institutes in Table 1 for kindly providing us bacterial species. The authors wish to thank to Pathology Laboratory, Department of Pathology, Faculty of Medicine, Srinakharinwirot University, HRH Princess Maha Chakri Sirindhorn Medical Center, Thailand for their participation in the measurement and analysis of the Bacteriological Analytical Manual (BAM) of the protocol of the Food and Drug Administration (FDA) standard method under the framework of this research. The authors also extend our appreciation to Mrs. Arda Pakpitchareon and Mr. Pisan Khawsak for technical assistance and Central Laboratory, Faculty of Medicine, Srinakharinwirot University, for scientific equipment.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Bangkok, 10110, Thailand

    Sirirat Wachiralurpan & Kosum Chansiri

  2. Faculty of Environmental Culture and Ecotourism, Srinakharinwirot University, Sukhumvit 23, Bangkok, 10110, Thailand

    Thayat Sriyapai

  3. Innovative Learning Center, Srinakharinwirot University, Sukhumvit 23, Bangkok, 10110, Thailand

    Supatra Areekit & Thongchai Kaewphinit

  4. Department of Biology, Faculty of Science, Srinakharinwirot University, Sukhumvit 23, Bangkok, 10110, Thailand

    Pichapak Sriyapai

  5. Department of Pathology, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Bangkok, 10110, Thailand

    Somchai Santiwatanakul

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  1. Sirirat Wachiralurpan
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  2. Thayat Sriyapai
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  3. Supatra Areekit
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  4. Thongchai Kaewphinit
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  6. Somchai Santiwatanakul
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  7. Kosum Chansiri
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Correspondence to Kosum Chansiri.

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Funding

This study was funded by Thailand Research Fund through the Royal Golden Jubilee Ph.D. Program (Grant No. PHD/0040/2554), The Graduate school of Srinakharinwirot University, and Agricultural Research Development Agency (Public organization), Thailand (No. CRP5705022280).

Conflict of Interest

Sirirat Wachiralurpan declares that she has no conflict of interest. Thayat Sriyapai declares that he has no conflict of interest. Supatra Areekit declares that she has no conflict of interest. Thongchai Kaewphinit declares that he has no conflict of interest. Pichapak Sriyapai declares that she has no conflict of interest. Somchai Santiwatanakul declares that he has no conflict of interest. Kosum Chansiri declares that she has no conflict of interest.

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Wachiralurpan, S., Sriyapai, T., Areekit, S. et al. Development of a Rapid Screening Test for Listeria monocytogenes in Raw Chicken Meat Using Loop-Mediated Isothermal Amplification (LAMP) and Lateral Flow Dipstick (LFD). Food Anal. Methods 10, 3763–3772 (2017). https://doi.org/10.1007/s12161-017-0949-4

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