Aptamer-conjugated silver nanoparticles for electrochemical dual-aptamer-based sandwich detection of staphylococcus aureus

Staphylococcus aureus (Staphylococcus aureus) is one of the most important human pathogens and causes numerous illnesses. In this study, we report a sensitive and highly selective dual-aptamer-based sandwich immunosensor for the detection of Staphylococcus aureus. In this bioassay system, a biotinylated primary anti-Staphylococcusaureus aptamer was immobilized on streptavidin coated magnetic beads (MB), which serves as a capture probe. A secondary anti-Staphylococcusaureus aptamer was conjugated to silver nanoparticles (Apt-AgNP) that sensitively reports the detection of the target. In the presence of target bacterium, an Apt/Staphylococcusaureus/apt-AgNP sandwich complex is formed on the MB surface and the electrochemical signal of AgNPs followed through anodic stripping voltammetry. The proposed sandwich assay benefits from advantageous of a sandwich assay for increased specificity, MB as carriers of affinity ligands for solution-phase recognition and fast magnetic separation, AgNPs for signal amplification, and an electrochemical stripping voltammetry read-out as a simple and sensitive detection. The electrochemical immunosensor shows an extended dynamic range from 10 to 1 x 10(6) cfu/mL with a low detection limit of 1.0 cfu/mL (S/N=3). Furthermore, the possible interference of other analog bacteria was studied. To assess the general applicability of this sensor, we investigated the quantification of Staphylococcus aureus in real water samples. The results were compared to the experimental results obtained from a plate counting method, which demonstrated an acceptable consistency. (C) 2014 Elsevier B.V. All rights reserved.