Scano-magneto immunoassay based on carbon nanotubes/gold nanoparticles nanocomposite for Salmonella enterica serovar Typhimurium detection

To improve sensitivity of S. enterica serovar Typhimurium detection, multiwalled carbon nanotubes (MWCNTs) and gold nanoparticles (AuNPs) were combined and used as a label to amplify signal in a scanometric based assay. In this study, the MWCNTs/AuNPs nanocomposite was fabricated by directly assemble of Au3+ to MWCNTs and allowed growing of AuNPs along the MWCNTs surface. This MWCNTs/AuNPs nanocomposite was then attached to anti-S. typhimurium antibody (MWCNTs/AuNPs/Ab1) and used as a detecting molecule. Upon binding to Salmonella, they were pre-concentrated by magenetic beads/antibody (MBs/Ab2) forming a sandwich immuno-complex which is later spotted on a nitrocellulose membrane coated slide. Silver reduction was applied to amplify signal. The detection limit of 42 CFU/ml was achieved when 2% BSA was used as a blocking agent. Given different types of real samples testing, chicken broth was found to give lowest detection limit, followed by orange juice low fat and whole milk. Selectivity testing was performed by using Escherichia coli as interference and found slightly cross-reactivity which could be due to specificity of the Ab used. By virtue of using a slide for multi-samples spotting and a flatbed scanner for signal-read out acquisition, this scano-magneto immunoassay could enable low-cost detection as well as high throughput screening.

► We enhance S. typhimurium detection sensitivity using MWCNTs/AuNPs nanocomposite. ► MWCNTs/AuNPs/Ab1 forms a complex with magnetic beads/Ab2 upon Salmonella detection. ► Signal is amplified through silver reduction of complex on nitrocellulose membrane. ► Signal-read out is simply acquired by a flatbed scanner. ► The detection limit of 42 CFU/ml was achieved when using this platform.