Abstract: In this article, two typical model strains of Cronobacter spp. (Cronobacter malonaticus and Enterobacter sakazakii) were used to establish a rapid detection system integrating one-step DNA extraction and rapid real-time polymerase chain reaction (real-time PCR) for Cronobacter spp.. The amplification time of the optimized fast real-time PCR was only 27 min 18 s compared to 82 min for the common procedure. in the one-step DNA extraction procedure, DNA could be directly obtained after thermal cyclic reaction for only 12 min. Compared with the traditional boiling method, the rapid detection method showed consistent sensitivity to pure culture and artificially contaminated samples cultured for 6 h. The sensitivity of the new method to artificially contaminated samples cultured for 7 and 8 h was one order of magnitude lower than that of the conditional method. The method presented in this study is fast, simple, and highly sensitive, and has a good application prospect in the rapid on-site detection of foodborne pathogens at ports.

Key words: microbial contamination; food safety; fast real-time polymerase chain reaction; Cronobacter spp.; quick detection technology