On-site monitoring of Escherichia coli O157:H7 in drinking water based on rapid detection of the rfbE gene at the single copy level

Abstract

The sensitive and on-site detection of highly pathogenic Escherichia coli O157:H7 (E. coli O157:H7) is of great importance to ensure public health. However, conventional methods for E. coli O157:H7 detection may suffer disadvantages such as high expense and time consumption. Herein, we reported a cost-effective colorimetric platform based on loop-mediated isothermal amplification (LAMP) for ultrasensitive and rapid detection of E. coli O157:H7. In this study, a novel quantitative colorimetric LAMP assay was established by in-situ measurement of hydroxy naphthol blue (HNB) color change with a designed portable device, which was applied to the amplification of the specific rfbE gene of E. coli O157:H7. As expected, this method featured a wide linear range (100-104 copies µL−1) with excellent linear regression analysis (R2 > 0.99) and possessed a record-low detection limit of 1 copy µL−1 to lower the risk of missed detection. The proposed strategy could be employed on drinking water samples with favorable recoveries and also demonstrated potential in the analysis of complex samples. This work exhibits the promising application prospect in on-site detection without the need for expensive instrumentation and seeks to provide an original paradigm for the rapid and sensitive detection of waterborne pathogens.