| Summary: | <i>Escherichia coli</i> are common pathogens, whereas <i>E. coli</i> O157:H7 is the most notorious <i>E. coli</i> strain, owing to its high virulence that can cause serious adverse effects and death. <i>E. coli</i> contains abundant peroxidases. Thus, the presence of <i>E. coli</i> can be determined by mixing <i>E. coli</i> with its substrate such as 3,5,3′,5′ tetramethylbenzidines (TMB) for endogenous peroxidase reactions. Under the presence of a high concentration of <i>E. coli</i>, colorless TMB turned to bluish, owing to the generation of the complexity of TMB and its oxidized TMB. To further reduce the detectable cell concentration, we developed an affinity-based method combined with an endogenous peroxidase reaction and mass spectrometric detection to detect <i>E. coli</i>. Affinity probes (diameter: ~20 µm) modified with maltose were generated for the enrichment of <i>E. coli</i> from sample solutions. <i>E. coli</i> trapped by the affinity probes was reacted with TMB in the presence of hydrogen peroxide for endogenous peroxidase reactions. Contactless atmospheric pressure ionization mass spectrometry was used for the detection of the reaction product, oxidized TMB (TMB cationic radical), to indicate the presence of target bacteria. The results showed that the developed method can be used to rapidly determine the presence of <i>E. coli</i> from a sample solution based on the detection of the TMB cationic radicals. The lowest detectable concentration of our method against <i>E. coli</i> O157:H7 in buffers and in complex juice samples was as low as ~100 cfu mL<sup>−1</sup>.
|
|---|