Multiplex pathogen detection based on spatially addressable microarrays of barcoded resins

Biotechnol J. 2008 Jul;3(7):948-53. doi: 10.1002/biot.200700236.

Abstract

Suspension microsphere immunoassays are rapidly gaining recognition in antigen identification and infectious disease biodetection due to their simplicity, versatility and high-throughput multiplex screening. We demonstrate a multiplex assay based on antibody-functionalized barcoded resins (BCRs) to identify pathogen antigens in complex biological fluids. The binding event of a particular antibody on given bead (fluorescence) and the identification of the specific pathogen agent (vibrational fingerprint of the bead) can be achieved in a dispersive Raman system by exciting the sample with two different laser lines. Anthrax protective antigen, Franciscella tularensis lipopolysaccharide and CD14 antigens were accurately identified and quantified in tetraplex assays with a detection limit of 1 ng/mL. The rapid, versatile and simple analysis enabled by the BCRs demonstrates their potential for multiplex antigen detection and identification in a reconfigurable microarray format.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Bacterial / isolation & purification*
  • Bacterial Toxins / isolation & purification*
  • Biosensing Techniques / methods
  • Fluoroimmunoassay / methods
  • Francisella tularensis / isolation & purification*
  • Humans
  • Lipopolysaccharide Receptors / isolation & purification
  • Lipopolysaccharides / isolation & purification
  • Microarray Analysis / methods*
  • Microspheres
  • Resins, Synthetic / chemistry*
  • Sensitivity and Specificity
  • Spectrum Analysis, Raman

Substances

  • Antigens, Bacterial
  • Bacterial Toxins
  • Lipopolysaccharide Receptors
  • Lipopolysaccharides
  • Resins, Synthetic
  • anthrax toxin