Development of an optical immunosensor based on the fluorescence of Cyanine-5 for veterinarian diagnostics.
A model optical immunosensor was developed to quantify an antibody present in a sample by measuring the fluorescence of Cyanine-5 conjugated with the antibody, using a competitive and a sandwich immunoreaction configuration, with the antigen immobilised in controlled pore glass beads. At pH 2, 94% of the antigen-antibody complex was dissociated, allowing reutilisation. Photobleaching had no effect on the fluorescence. This model system was used to detect Brucella sp. infection and could quantify anti-Brucella sp. antibodies in ovine serum samples in the range from 0.005 to 0.11 mg ml(-1).
Biosensors Laboratory, IBET/ITQB- Instituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Química e Biológica, Apartado 12, P-2781-901 Oeiras, Portugal.
Marta
M
Silva
Helder
H
Cruz
Osvaldo
O
Rossetti
Alicia
A
Arese
Abel
A
Oliva
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods / Q000662:veterinary*; D002002:Brucella / Q000276:immunology* / Q000302:isolation & purification*; D002007:Brucellosis, Bovine / Q000097:blood / Q000175:diagnosis / Q000382:microbiology; D002232:Carbocyanines; D002417:Cattle; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005456:Fluorescent Dyes; D015200:Fluoroimmunoassay / Q000295:instrumentation* / Q000379:methods / Q000662:veterinary*; D008565:Membrane Proteins / Q000276:immunology*; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods; D055095:Optics and Photonics / Q000295:instrumentation; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012756:Sheep; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods / Q000662:veterinary
C106040:BP26 protein, Brucella; D002232:Carbocyanines; D005456:Fluorescent Dyes; D008565:Membrane Proteins; C085321:cyanine dye 5