Combining phagomagnetic separation with immunoassay for specific, fast and sensitive detection of Staphylococcus...

A Staphylococcus aureus (S. aureus)-specific lytic bacteriophage P-S. aureus-9, isolated from an environmental water sample, was assembled on magnetic beads for capturing S. aureus from samples through magnetic separation. Horseradish Peroxidase (HRP) labeled immunoglobulin (IgG) antibodies were used to detect the captured S. aureus by reacting with protein A on S. aureus followed by colorimetric signals, which were generated from the catalytic reaction between HRP and the substrate 3,3',5,5'-Tetramethylbenzidine (TMB). Under optimal conditions, the calibration curve was linear from 1.0×10⁴ to 1.0×10⁶CFUmL^-1. The limit of detection (LOD) for the assay was 2.47×10³CFUmL^-1 and 8.86×10³CFUmL^-1 of S. aureus in PBS and apple juice, respectively. Moreover, the whole assay revealed outstanding specificity towards S. aureus, without any interference of common pathogenic bacteria, and can be completed within 90min without any pre-enrichment. As far as known, it was the first time to detect S. aureus based on the double site recognition of bacteriophage and mammal IgG. The novel approach has shown good potentials for a rapid, specific, cheap and simple detection of S. aureus in food samples.