Escherichia coli O157:H7, a major foodborne pathogen, is a major public health concern associated with life-threatening diseases such as hemolytic uremic syndrome. To alleviate this burden, a sensitive and rapid system is required to detect this pathogen in various kinds of foods. Herein, we propose a phage-based pathogen detection method to replace laborious and time-consuming conventional methods. We engineered an E. coli O157:H7-specific phage phiV10 to rapidly and sensitively detect this notorious pathogen. The luxCDABE operon was introduced into the phiV10 genome and allowed the engineered phage phiV10lux to generate bioluminescence proportional to the number of viable E. coli O157:H7 cells without any substrate addition. The phage phiV10lux was able to detect at least 1 CFU/ml of E. coli O157:H7 in a pure culture within 40 min after 5 h of pre-incubation. In artificially contaminated romaine lettuce, apple juice (pH 3.51), and ground beef, the reporter phage could detect approximately 10 CFU/cm2, 13 CFU/ml, and 17 CFU/g of E. coli O157:H7, respectively. Taken together, the constructed reporter phage phiV10lux could be applied as a powerful tool for rapid and sensitive detection of live E. coli O157:H7 in foods.
Bibliographical noteFunding Information:
We would like to thank Dr. Rafid Ahmed at the National Microbiology Laboratory, Canadian Science Center for Human and Animal Health (Winnipeg, Canada), for kindly providing of phage phiV10 and E. coli O157:H7 OE50. We also thank Prof. Kang at the Food Safety Engineering Laboratory, Seoul National University (Seoul, Korea), for providing the E. coli O157:H7 strains used in this experiment (O157:H7 2321, O157:H7 2324, O157:H7 3041, O157:H7 3046). This work was supported by Basic Science Research Programs (2016R1C1B2015578 and 2014R1A2A1A10051563) through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning.
All Science Journal Classification (ASJC) codes
- Food Science