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Detection of Pseudomonas fluorescens and Related Psychrotrophic Bacteria in Refrigerated Meat by a Sandwich ELISA
I González1, R Martín1, T García1
1Departamento de Nutrición y Bromatología III, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain.
Abstract:
A sandwich enzyme-linked immunosorbent assay (ELISA) was developed for detection of Pseudomonas fluorescens and related psychrotrophic bacteria in refrigerated meat. Polyclonal antibodies were raised in rabbits against protein F from the cell envelope of P. fluorescens AH-70. The ELISA involved capturing antigens from the microorganisms present on meat samples with the anti-protein F antibodies immovilized on 96-well plates, and detecting bound antigens with the same anti-PF antibodies conjugated to biotin. Commercial ExtrAvidin-peroxidase conjugate was used to detect the biotinylated antibodies bound to their specific antigens. Subsequent enzymatic conversion of substrate gave distinct absorbance differences when assaying meat samples containing P. fluorescens strains of different origin as well as related psychrotrophic microorganisms. The detection threshold for the ELISA assay developed in this work is 105 CFU/cm2.
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