Abstract
A simple, rapid, and sensitive electrochemical biosensor based on a screen-printed carbon electrode (SPCE) was developed for onsite detection of E. coli in real time. This work analyzed the effect of aptamer conjugation and PBS buffer solution on the colloidal stability of the silver nanoparticles (AgNPs). Aggregations of the AgNPs after aptamer conjugation in PBS buffer were observed from the particle size distribution analysis. The AgNP-aptamer conjugation and its affinity towards E. coli (DH5α) were confirmed by UV-visible spectrophotometry, which showed a linear increment in the absorption with increasing E.coli concentration. The screen-printed carbon electrodes were modified by drop-casting of AgNPs, which were used as an effective immobilization platform for E. coli-specific aptamers. The modified electrode's surface modification and redox behavior were characterized using cyclic voltammetry. Finally, E. coli was detected using differential pulse voltammetry with an optimized incubation time of 15 min. The developed biosensors showed a linear decrease in current intensity with an increase in the concentration of E. coli. The biosensor had a relative standard deviation (RSD) of 6.91% (n = 3), which showed good reproducibility. The developed biosensors are highly sensitive and have a limit of detection (LOD) as low as 150 CFU/ml. The biosensor showed good selectivity for E.coli coli when comparing the signal response obtained for bacteria other than E.coli. Also, the biosensor was found stable for four weeks at room temperature and showed high recoveries from 95.27% to 107% during the tap water sensitivity validation.