Immunoassay for foodborne pathogenic bacteria using magnetic composites Ab@Fe3O4, signal composites Ap@PtNp, and thermometer readings

Abstract

A point-of-care (POC) immunoassay was established for the sensitive and rapid detection of pathogenic Escherichia coli O157:H7, using magnetic Fe₃O₄ organic-inorganic composites (Ab@Fe₃O₄) for immunomagnetic separation, nanozyme platinum nanoparticle (PtNp) organic-inorganic composites (Ap@PtNp) for signal amplification, and thermometer readings. Antibodies and Fe₃O₄ were incubated in Cu²⁺ phosphate buffer to synthesize the magnetic composite Ab@Fe₃O₄ with antibodies, to specifically capture E. coli O157:H7. Antimicrobial peptides and PtNp were incubated in Cu²⁺ phosphate buffer to synthesize the signal composites Ap@PtNp with antimicrobial peptides (magainin I), recognizing and labeling E. coli O157:H7. In the presence of E. coli O157:H7, magnetic microcomposites targeted bacteria and signal microcomposites to form the sandwich structure: Ab@Fe₃O₄-bacteria-Ap@PtNp for magnetic separation. Ap@PtNp of signal composites catalyzed H₂O₂ to generate thermo-signals (temperature rise), which were determined by a thermometer. This point-of-care bioassay detected E. coli O157:H7 in the linear range of 10¹–10⁷ CFU mL⁻¹ and with a detection limit of 14 CFU mL⁻¹. One-pot process magnetic Fe₃O₄ organic-inorganic composites (Ab@Fe₃O₄, magnetic microcomposites, MMC) for immunomagnetic separation and nanozyme platinum nanoparticle (PtNp) organic-inorganic composites (Ap@PtNp, signal microcomposites, SMC) were used as signal amplification and thermometer readings for E. coli O157:H7 detection.