Colorimetric immunoassay for rapid detection of Staphylococcus aureus based on etching-enhanced peroxidase-like...

A novel colorimetric immunoassay for the detection of Staphylococcus aureus (S. aureus) based on a combination of immunomagnetic separation and signal amplification via etching-enhanced peroxidase-like catalytic activity of gold nanoparticles (AuNPs) was developed. Nanoconjugates composed of gold and iron oxide nanoparticles were synthesized and further modified with antiS. aureus immunoglobulin Y (IgY), which was used for the selective enrichment and rapid separation of target bacteria in complex matrices. AuNPs functionalized with antiS. aureus aptamer were used as an artificial enzyme which has peroxidase-like catalysis activity. Catalytic activity of AuNPs is inhibited by modifying aptamer. However, catalysis of modified AuNPs remarkably enhanced by hydrogen peroxide etching. Based on collecting unbound modified AuNPs in the supernatant and 3,3',5,5'-tetramethylbenzidine-hydrogen peroxide reporting system, the yellow color of solution decreases linearly with increasing the concentration of S. aureus ranging from 10 to 10⁶ cfu/mL. The limit of detection is 10 cfu/mL, and total detection time is 65 min. The recoveries of the S. aureus spiked in food samples are 88.2-119.8%. Schematic illustration of colorimetric method for detection of S. aureus based on the IgY-Fe₃O₄/Au nanocomposites as capture probes and apt-AuNPs as artificial enzyme with etching-enhanced peroxidase-like catalytic activity.