Microchimica Acta

, 163:203

Detection of pathogenic Staphylococcus aureus bacteria by gold based immunosensors

  • Souhir Boujday
  • Romain Briandet
  • Michèle Salmain
  • Jean-Marie Herry
  • Pierre-Guy Marnet
  • Michel Gautier
  • Claire-Marie Pradier
Original Paper

DOI: 10.1007/s00604-008-0024-3

Cite this article as:
Boujday, S., Briandet, R., Salmain, M. et al. Microchim Acta (2008) 163: 203. doi:10.1007/s00604-008-0024-3

Abstract

We describe the elaboration of ultra-sensitive immunosensors, to detect the bacterial pathogen Staphylococcus aureus. We utilized commercially available polyclonal anti-S. aureus antibody as receptor. Immunosensors were elaborated by building a self-assembled monolayer (SAM) of thiolamine onto planar gold-coated sensor chips. Then, Protein A was covalently linked to the thiolated SAM using glutaraldehyde as cross-linking agent. After a blocking step by Bovine Serum Albumin (BSA), the antibody was immobilized by affinity to Protein A. This step-by-step construction was monitored by Polarization Modulation Reflection Absorption Infrared Spectroscopy (PM-RAIRS) and Quartz Crystal Microbalance with Dissipation (QCM-D). In a first stage, the parameters of immunosensor elaboration were optimized using a model rabbit IgG. The accessibility of receptors and the homogeneity of their distribution were checked by PM-RAIRS, QCM-D, and by immuno-gold scanning electron microscopy. Then, the specific rabbit anti-S. aureus antibody was immobilized and the resulting sensing layer was applied to the detection of the pathogen target. Independent detection of bacteria immobilized on the sensors by fluorescent imaging allowed validation of the specificity of recognition toward the pathogen as well as a quantitative response of the sensor. Using PM-RAIRS as transducing technique allowed us to enhance sensitivity and reach a very competitive detection level (105 CFU mL−1).

Keywords

ImmunosensorsPM-RAIRSQCM-DSEMStaphylococcus aureus

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Souhir Boujday
    • 1
    • 2
  • Romain Briandet
    • 3
  • Michèle Salmain
    • 4
  • Jean-Marie Herry
    • 3
  • Pierre-Guy Marnet
    • 5
  • Michel Gautier
    • 5
  • Claire-Marie Pradier
    • 1
    • 2
  1. 1.UPMC Univ Paris 6, UMR CNRS 7609Laboratoire de Réactivité de SurfaceParisFrance
  2. 2.CNRS, UMR 7609Laboratoire de Réactivité de SurfaceParisFrance
  3. 3.UMR 763 INRA-AgroParisTechBioadhésion et Hygiène des MatériauxMassyFrance
  4. 4.Laboratoire de Chimie et Biochimie des Complexes Moléculaires, UMR CNRS 7576Ecole Nationale Supérieure de Chimie de ParisParisFrance
  5. 5.Laboratoire de microbiologieUMR 1263 INRA-AgrocampusRennes cedexFrance