Label-Free Detection of Staphylococcus aureus Captured on Immutable Ligand Arrays

The rapid capture and label-free detection of Staphylococcus aureus , an opportunistic bacterium that can infect upon contact, can be performed using periodic microarrays of ligand-protein conjugates created by noncontact (inkjet) printing, darkfield imaging conditions, and a FFT-based readout method. Ink solutes were prepared using bovine serum albumin (BSA) conjugated to a glycan with high affinity for bacterial adhesins and printed as dot-matrix arrays with periodicities of 80-120 μm using a thermal injection method. Upon exposing the glycan-BSA microarrays to live strains of S. aureus , patterns emerge that can be detected under optical darkfield conditions. These patterns can be decoded by fast Fourier transform (FFT) analysis to generate fault-tolerant readout signals that correspond to the capture of S. aureus, with a limit of detection between 10(2) and 10(3) cfu/mL. Inkjet printing provides independent control over array periodicity, enabling FFT signals to be assigned to specific frequencies in reciprocal k-space.