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

Yufei Chen, Hao Li, Liu Yang, Lei Wang, Ruyi Sun, Julia E. S. Shearer and Fengjie Sun
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Yufei Chen: School of Grain Science and Technology, Jilin Business and Technology College, Changchun 130507, China
Hao Li: College of Food Engineering, Jilin Engineering Normal University, Changchun 130052, China
Liu Yang: School of Grain Science and Technology, Jilin Business and Technology College, Changchun 130507, China
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

IJERPH, 2021, vol. 18, issue 9, 1-16

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.

Keywords: Clostridium botulinum; botulinum neurotoxin; LAMP; ntnh; turbidity method; fluorescence method (search for similar items in EconPapers)
JEL-codes: I I1 I3 Q Q5 (search for similar items in EconPapers)
Date: 2021
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