Abstract: To develop a rapid detection method for Clostridium perfringens using real-time polymerase chain reaction (PCR) or real-time recombinase polymerase amplification (RPA), we designed specific primers and probe based on the conserved sequence of the plc gene of C. perfringens. The results of specificity analysis showed that the two established methods specifically detected C. perfringens but not other bacteria. The sensitivity of both methods was 1.3 pg/μL and the limit of detection for C. perfringens in artificially contaminated chicken and milk samples was 1.0 × 102 CFU/mL. The positive samples could be detected in 3–13 min by real-time RPA, and at least 24–46 min by real-time PCR (Ct = 17.45–33.65). Real-time RPA was better than real-time PCR with respect to analysis time, easiness of operation and portability. In conclusion, thanks to their high specificity and sensitivity and easy operation, real-time RPA and real-time PCR provided an effective technical approach for the rapid detection of C. perfringens.

Key words: Clostridium perfringens, plc gene, real-time recombinase polymerase amplification, real-time polymerase chain reaction