RESUMO
The precise detection of pathogenic microorganisms is crucial for the reduction of water-borne diseases. Herein, a filter-paper-based florescent chemosensor was fabricated for the detection of Escherichia coli and Staphylococcus aureus contamination exploiting protein-DNA interaction between the target and a specific probe. The sensing mechanism involved the self-assembly of Rhodamine B (RhB) on silver nanoparticles (AgNPs) surface that was labeled with a single-stranded DNA probe. This causes the fluorescence quenching of RhB by a distant-dependant process. The hybridization between pathogen-specific probe and bacterial surface protein causes the release of fluorescence of RhB, which was observed under UV light. For paper-based bio-surface preparation, the mixture comprising RhB-AgNP-ssDNA was drop-casted on filter paper discs. The conditions were optimized using isolated genomic DNA of the microbes. The method was applied for E.coli detection using an eae gene-based probe targeting intimin protein and S. aureus detection using tuf gene-based probe targeting EF-tuf protein on the microbe's surface. The chemosensor had a notable specificity and selectivity for E.coli, and S. aureus, with detection limits of 0.6 × 108 and 0.37 × 103 CFU/mL respectively. Moreover, the sensor was tested on real water samples, which presented excellent reproducibility of results (RSD ≤ 0.24%). Furthermore, the gradient change of fluorescence was captured by a smartphone, which allows direct detection of pathogens in a sensitive semi-quantitative way without the need for expensive instruments. The designed chemosensor can serve as a simple, inexpensive, and rapid method for the on-site detection of microbial contamination in drinking water.