Multiplex pathogen detection based on spatially addressable microarrays of barcoded resins.
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.
Blais
David R
DR
The Steacie Institute for Molecular Sciences, National Research Council of Canada, Ottawa, Canada.
Alvarez-Puebla
Ramon A
RA
Bravo-Vasquez
Juan P
JP
Fenniri
Hicham
H
Pezacki
John Paul
JP
D000942:Antigens, Bacterial / Q000302:isolation & purification*; D001427:Bacterial Toxins / Q000302:isolation & purification*; D015374:Biosensing Techniques / Q000379:methods; D015200:Fluoroimmunoassay / Q000379:methods; D005604:Francisella tularensis / Q000302:isolation & purification*; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000302:isolation & purification; D008070:Lipopolysaccharides / Q000302:isolation & purification; D046228:Microarray Analysis / Q000379:methods*; D008863:Microspheres; D012117:Resins, Synthetic / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013059:Spectrum Analysis, Raman
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D012117:Resins, Synthetic; C030325:anthrax toxin