Colorimetric immunoassay for Listeria monocytogenes by using core gold nanoparticles, silver nanoclusters as oxidase mimetics, and aptamer-conjugated magnetic nanoparticles

Abstract

The authors describe a rapid colorimetric assay for Listeria monocytogenes (L. monocytogenes) based on the o-phenylenediamine-mediated deaggregation of gold nanoparticles. Silver nanoclusters are used as an artificial enzyme that can oxidize o-phenylenediamine to form o-benzoquinone diamine. Aptamer and IgY antibodies were chosen to conjugate with magnetic beads and silver nanoclusters, respectively, which can recognize and bind L. monocytogenes at different specific binding sites. This results in the disassembly of colloidal gold nanoparticles which is accompanied by a color change from blue to red, with peaks at 730 and 525 nm, respectively. The method allows L. monocytogenes to be colorimetrically determined in the 10 to 10⁶ cfu·mL^-1 concentration range without pre-enrichment, and the limit of detection is as low as 10 cfu·mL^-1. Recoveries ranging from 97.4 to 101.3% are found when analyzing spiked food samples. The assay is rapid, sensitive and specific. Graphical abstract Schematic illustration of a colorimetric method for detection of L. monocytogenes based on silver nanoclusters-catalyzed oxidation of OPD and de-aggregation of GNPs. A color change from blue to red can be observed and correlated to the concentration of L. monocytogenes.

Keywords: Food safety; Immunomagnetic separation; Nanozyme; Visual detection; de-aggregation; o-phenylenediamine.