Development and application of the loop‐mediated isothermal amplification assay for rapid detection of enterotoxigenic Clostridium perfringens in food

Abstract

AbstractA loop‐mediated isothermal amplification (LAMP) method for rapid detection of enterotoxigenic Clostridium perfringens in food is developed and evaluated in this study. Six primers were designed to recognize the cpa gene of C. perfringens. A panel of 45 bacterial strains, including 15 C. perfringens and 30 other strains, were included in this study to evaluate and optimize the LAMP assay. The specificity of the LAMP assay was 100%. The sensitivity of the LAMP assay for the detection of C. perfringens in food was 10 CFU/ml. Different methods of extracting DNA from artificially contaminated food, such as boiling, NaOH treatment, and the use of magnetic beads, were evaluated. The established LAMP assay was used to analyze C. perfringens in various food samples. The magnetic bead‐based method was a useful tool for extracting genomic DNA from 14 foods contaminated by C. perfringens. All 15 foods contaminated by C. perfringens were identified as positive.Practical ApplicationsThe traditional method for detecting Clostridium perfringens in foods involves the use of a bioassay that is laborious and time‐consuming. The LAMP assay is a useful and powerful tool for the rapid detection of C. perfringens, and, undoubtedly, the efficiency, technical simplicity, and cost‐effectiveness of the LAMP assay will have broad applications in bacteriological detection of enterotoxigenic C. perfringens.