Sensitive and rapid amperometric magnetoimmunosensor for the determination of Staphylococcus aureus

Anal Bioanal Chem. 2012 May;403(4):917-25. doi: 10.1007/s00216-012-5738-8.

Abstract

The preparation and characteristics of a disposable amperometric magnetoimmunosensor, based on the use of functionalized magnetic beads (MBs) and gold screen-printed electrodes (Au/SPEs), for the specific detection and quantification of Staphylococcal protein A (ProtA) and Staphylococcus aureus (S. aureus) is reported. An antiProtA antibody was immobilized onto ProtA-modified MBs, and a competitive immunoassay involving ProtA antigen labelled with HRP was performed. The resulting modified MBs were captured by a magnetic field on the surface of tetrathiafulvalene-modified Au/SPEs and the amperometric response obtained at -0.15 V vs the silver pseudo-reference electrode of the Au/SPEs after the addition of H2O2 was used as transduction signal. The developed methodology showed very low limits of detection (1 cfu S. aureus/mL of raw milk samples), and a good selectivity against the most commonly involved foodborne pathogens originating from milk. These features, together with a short analysis time (2 h), the simplicity, and easy automation and miniaturization of the required instrumentation make the developed methodology a promising alternative in the development of devices for on-site analysis.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / immunology
  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods*
  • Cattle
  • Humans
  • Immunoassay / instrumentation
  • Immunoassay / methods*
  • Magnetics / methods*
  • Milk / chemistry
  • Milk / microbiology
  • Sensitivity and Specificity
  • Staphylococcal Infections / diagnosis
  • Staphylococcal Infections / microbiology*
  • Staphylococcal Protein A / immunology
  • Staphylococcus aureus / immunology
  • Staphylococcus aureus / isolation & purification*

Substances

  • Antibodies, Bacterial
  • Staphylococcal Protein A