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Rapid Detection of Clostridium botulinum in Food Using Loop-Mediated Isothermal Amplification (LAMP)

Author

Listed:
  • Yufei Chen

    (School of Grain Science and Technology, Jilin Business and Technology College, Changchun 130507, China
    These authors contributed equally to this work.)

  • Hao Li

    (College of Food Engineering, Jilin Engineering Normal University, Changchun 130052, China
    These authors contributed equally to this work.)

  • Liu Yang

    (School of Grain Science and Technology, Jilin Business and Technology College, Changchun 130507, China
    These authors contributed equally to this work.)

  • Lei Wang

    (School of Grain Science and Technology, Jilin Business and Technology College, Changchun 130507, China)

  • Ruyi Sun

    (College of Life Sciences, Jilin Agricultural University, Changchun 130118, China)

  • Julia E. S. Shearer

    (School of Science and Technology, Georgia Gwinnett College, Lawrenceville, GA 30043, USA)

  • Fengjie Sun

    (School of Science and Technology, Georgia Gwinnett College, Lawrenceville, GA 30043, USA)

Abstract

Botulinum neurotoxins are considered as one of the most potent toxins and are produced by Clostridium botulinum . It is crucial to have a rapid and sensitive method to detect the bacterium Clostridium botulinum in food. In this study, a rapid detection assay of C. botulinum in food using loop-mediated isothermal amplification (LAMP) technology was developed. The optimal primers were identified among three sets of primers designed specifically based on the partial ntnh gene encoding nontoxic-nonhaemagglutinin (NTNH) for rapid detection of the target DNA in plasmids. The optimal temperature and reaction time of the LAMP assay were determined to be 64 °C and 60 min, respectively. The chemical kit could be assembled based on these optimized reaction conditions for quick, initial high-throughput screening of C. botulinum in food samples. The established LAMP assay showed high specificity and sensitivity in detecting the target DNA with a limit of 0.0001 pg/ul (i.e., ten times more sensitive than that of the PCR method) and an accuracy rate of 100%. This study demonstrated a potentially rapid, cost-effective, and easy-operating method to detect C. botulinum in food and clinical samples based on LAMP technology.

Suggested Citation

  • Yufei Chen & Hao Li & Liu Yang & Lei Wang & Ruyi Sun & Julia E. S. Shearer & Fengjie Sun, 2021. "Rapid Detection of Clostridium botulinum in Food Using Loop-Mediated Isothermal Amplification (LAMP)," IJERPH, MDPI, vol. 18(9), pages 1-16, April.
    • Handle: RePEc:gam:jijerp:v:18:y:2021:i:9:p:4401-:d:540241
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