Optimization of Bacterial Concentration by Filtration for Rapid Detection of Foodborne Escherichia coli O157:H7 Using Real‐Time PCR Without Microbial Culture Enrichment

Abstract Escherichia coli O157:H7 is an important foodborne pathogen and has been implicated in numerous food poisoning outbreaks worldwide. Although several microbiological and molecular methods have been developed to detect E. coli O157:H7, the difficulty to rapidly detect low levels of the foodborne bacteria persists. Here, the optimization of a filtration technique to concentrate and rapidly detect E. coli O157:H7 was conducted. Using homogenates prepared from freshly cut lettuce and cabbage samples, the E. coli O157:H7 concentration efficiencies of seven membrane filters were compared. Mixed cellulose ester (MCE) and polyvinylidene difluoride (PVDF) filters demonstrated the highest bacterial recoveries. In addition, the optimal E. coli O157:H7 detachment method from MCE filters after filtration was investigated. Tapping for 80 s was demonstrated to be the most effective method for detaching bacteria from the filters. Further, the possibility of the rapid detection of low levels of E. coli O157:H7 in lettuce and cabbage was evaluated using real‐time polymerase chain reaction after bacterial concentration using MCE and PVDF filters. The use of MCE filters enabled the detection of 10° CFU/g (5 CFU/g) of E. coli O157:H7 within 2 hr without microbial enrichment culture. Therefore, concentration by filtration can be used for the rapid detection of low levels of foodborne pathogens. Practical Application The modified method, which has been verified in this study, has been optimized to reduce the analysis time and to detect very low concentrations of E. coli O157:H7 within 2 hr. All these detection systems have a direct economic impact on the food analysis of producers, health authorities, or third‐party laboratories.