Rapid detection Clostridium perfringens by recombinase polymerase ampification (RPA)

Abstract

    Clostridium perfringens has been considered as one of pathogens contributing to foodborne illness ranking only after Salmonella. C. perfringens is a Gram-positive, anaerobic bacterium, which can be abundantly found in the wide range of places such as soil, animal intestine and frozen meat. C. perfringens causes food poisoning in human with stomachache and diarrhea. In order to detect this pathogenic bacterium, several isolated, cultured, biochemical methods as well as PCR technique could be performed under complicated processes which is time-consuming. This also requires experienced technicians and an expensive thermal cycler. This study applies RPA (recombinase polymerase amplification) isothermal nucleic acid amplification technique to rapidly detect Clostridium perfringens. RPA has a short reaction time for target DNA replication, is performed at only one temperature, and achieves high sensitivity and specificity. The results have established an accurate detection procedure characteristic of C. perfringens in 25 minutes at 38 ⁰C, 10 times more sensitive than PCR techniques, creating a premise for direct detection. C. perfringens on food samples.