The fluorescent probe-based recombinase-aided amplification for rapid detection of Escherichia coli O157:H7 |
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| Affiliation: | 1. Hebei Medical University, Shijiazhuang 050031, Hebei, China;2. Hebei General Hospital, Shijiazhuang 050070, Hebei, China;3. NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;4. State Key Laboratory of Infectious Disease Prevention and Control, National Institute forCommunicable DiseaseControl and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;5. Wuxi Center for Disease Control and Prevention, Wuxi 214023, Jiangsu, China;6. Shandong Provincial Research Center for Bioinformatic Engineering and Technique, School of Life Sciences and Medicine, Shandong University of Technology, Zibo255049, Shandong, China;7. Laboratory of Virology, Beijing Key Laboratory ofEtiology of Viral Diseases in Children, Capital Institute of Pediatrics, Beijing 100020, China |
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| Abstract: | Escherichia coli O157:H7 (E. coli O157:H7) is a common foodborne morbigenous microorganism, which can spread through fecal-oral transmission. Humans can be infected by ingesting foods and water contaminated with E. coli O157:H7, which can cause various symptoms. In present study, we have successfully developed a quick and hypersensitive fluorescent probe-based Recombinase-aided amplification (RAA) method and applied in E. coli O157:H7 detection at 39 °C in 20 min. The sensitivity of the assay in pure E. coli O157:H7 suspension was 5.6 × 100 CFU/mL. The fluorescent probe-based RAA assay was further applied in three samples, and the limit of detection (LOD) in skimmed milk, lettuces and lake water was 5.4 × 101 CFU/mL, 7.9 × 101 CFU/mL and 5.2 × 101 CFU/mL, separately. This method showed a high sensitivity and short detection time, which has the feasible application in on-site test in real samples. |
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| Keywords: | Recombinase-aided amplification Isothermal amplification Applications |
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