Signal-off colorimetric and signal-on fluorometric dual-mode aptasensor for ultrasensitive detection of Salmonella Typhimurium using graphitic carbon nitride

Food safety is severely burdened by the prevalence of foodborne pathogens and the diseases they cause, necessitating the development of rapid, easy-to-use, highly sensitive, and reliable detection methods. Here, a signal-off colorimetric and signal-on fluorometric dual-mode detection method for Salmonella Typhimurium (S. typhimurium) was developed based on its unique interaction with aptamer DNA and graphitic carbon nitride (GCN). In the absence of a target Salmonella species, 6-carboxyfluorescein (FAM)-labeled aptamers are adsorbed on the surface of GCN primarily via a π-π interaction, resulting in reduced fluorescence of FAM through GCN-mediated quenching as well as improved peroxidase-like activity of GCN to generate intense blue color through facilitated electrostatic attraction between the negatively charged aptamer and positively charged 3,3',5,5'-tetramethylbenzidine (TMB) substrate. The introduction of S. typhimurium to the sample solution causes the detachment of the aptamer from GCN due to its higher affinity for S. typhimurium than GCN, thereby rapidly reducing the colorimetric signal and recovering the fluorescence. We successfully determined the number of S. typhimurium using this method in a remarkably short duration (10-30 min), highlighting its rapidity. The limit of detection values for S. typhimurium were as low as 8 and 3 CFU/mL when using colorimetric and fluorometric methods, respectively. Moreover, this method can be used to detect S. typhimurium spiked in real vegetable extract and milk with high reproducibility and reliability. This method serves as a convenient route to the rapid, sensitive, selective, and reliable detection of pathogens from complex food samples, with the potential to replace conventional yet laborious methods currently in use.