Abstract
Rapid detection of food-borne pathogens is essential to public health and the food industry. Although the conventional culture method is highly sensitive, it takes at least a few days to detect food-borne pathogens. Even though polymerase chain reaction (PCR) can detect food-borne pathogens in a few hours, it is more expensive and unsatisfactorily sensitive relative to the culture method. We have developed a method to rapidly detect Salmonella enterica by using a compact disc (CD)-shaped device that can reduce reagent consumption in conventional PCR. The detection method, which combines culture and PCR, is more rapid than the conventional culture method and is more sensitive and cheaper than PCR. In this study, we also examined a sample preparation method that involved collecting bacterial cells from food. The bacteria collected from chicken meat spiked with S. enterica were mixed with PCR reagents, and PCR was performed on the device. At a low concentration of S. enterica, the collected S. enterica was cultured before PCR for sensitive detection. After cultivation for 4 h, S. enterica at 1.7 × 104 colony-forming units (CFUs)·g−1 was detected within 8 h, which included the time needed for sample preparation and detection. Furthermore, the detection of 30 CFUs·g−1 of S. enterica was possible within 12 h including 8 h for cultivation.
Highlights
Rapid detection of food-borne pathogens is of great importance to public health and the food industry
E. colifrom was aused as the general bacteria, because bacteria
−1 bacteria for non-heated food was less than 10 colony-forming units (CFUs)·g
Summary
Rapid detection of food-borne pathogens is of great importance to public health and the food industry. The PCR mixture contained DNA was divided into eleven thousands 1.8 nL droplets and PCR on the disk was performed. In this device, only 1 μL of PCR reagent is needed for one sample, so the amount of reagents consumed in our device is 25 times lower than that for conventional PCR, i.e., 25 μL. We evaluated the speed of detection of S. enterica in food using a combination of culture and PCR to improve the LOD of S. enterica in the device. By combining the culture method and PCR on the CD-shaped device, our detection method was more rapid than the conventional culture method, and more sensitive than real-time PCR based method
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