Abstract
Clostridium perfringens is a key anaerobic pathogen causing food poisoning. Definitive detection by standard culture method is time-consuming and labor intensive. Current rapid commercial test kits are prohibitively expensive. It is thus necessary to develop rapid and cost-effective detection tool. Here, loop-mediated isothermal amplification (LAMP) in combination with a lateral-flow biosensor (LFB) was developed for visual inspection of C. perfringens-specific cpa gene. The specificity of the developed test was evaluated against 40 C. perfringens and 35 other bacterial strains, which showed no cross-reactivity, indicating 100% inclusivity and exclusivity. LAMP-LFB detection limit for artificially contaminated samples after enrichment for 16 h was 1–10 CFU/g sample, which was comparable to the commercial real-time PCR kit. The detection performance of LAMP-LFB was also compared to culture-based method using 95 food samples, which revealed the sensitivity (SE), specificity (SP) and Cohen's kappa coefficient (κ) of 88.0% (95% CI, 75.6%-95.4%), 95.5% (95% CI, 84.8%-99.4%) and 0.832 (95% CI, 0.721–0.943), respectively. Area under the receiver operating characteristic (ROC) curve was 0.918 (95% CI, 0.854–0.981), indicating LAMP-LFB as high relative accuracy test. In conclusion, LAMP-LFB assay is a low-cost qualitative method and easily available for routine detection of C. perfringens in food samples, which could serve as an alternative to commercial test kit.
Highlights
Clostridium perfringens is as an anaerobe responsible for a wide range of diseases such as gas gangrene and food poisoning in human, necrotic enteritis in animals worldwide [1]
The positive results of loop-mediated isothermal amplification (LAMP)-lateral-flow biosensor (LFB) were in accordance with those obtained from the gel electrophoresis
The novel LAMP-LFB assay was successfully developed to detect of C. perfringens in food samples
Summary
Clostridium perfringens is as an anaerobe responsible for a wide range of diseases such as gas gangrene and food poisoning in human, necrotic enteritis in animals worldwide [1] It is one of the most common pathogens that causes foodborne illness in the United States. Rapid diagnostic tests based on molecular detection such as real-time PCR have been commercially available for detection of foodborne pathogens including C. perfringens [7]. Such detection kit represents qualitative detection of a highly conserved DNA sequence specific to the cpa gene, which is presented in all C. perfringens strains [8,9,10]. It is of interest to develop a rapid detection tool, which is cost-effective, sensitive, labor-saving and exhibits immediate results as an alternative to the current methods
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