Fluorescence polarization-based homogeneous assay for...

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G01N 33/58 (2006.01) G01N 33/533 (2006.01) G01N 33/542 (2006.01)

Patent

CA 2452493

A homogenous assay for determining the aflatoxin content in agricultural products uses the technique of fluorescence polarization. A solvent is used to extract aflatoxins from a sample of the agricultural product. A mixture is prepared by combining the extract with a tracer and with a monoclonal antibody specific for aflatoxin. The tracer is able to bind to the monoclonal antibody to produce a detectable change in fluorescence polarization. The tracer is prepared by conjugating an aflatoxin oxime to a suitable fluorophore. The fluorescence polarization of the mixture is measured. The aflatoxin concentration of the mixture may be calculated using a standard curve obtained by measuring the fluorescence polarization of a series of aflatoxin solutions of known concentration.

Un dosage homogène permettant de déterminer la teneur en aflatoxines de produits agricoles utilise la technique de la polarisation de fluorescence. Pour extraire les aflatoxines présentes dans un échantillon du produit agricole on utilise un solvant. On prépare un mélange formé par la combinaison de l'extrait avec un traceur et avec un anticorps monoclonal spécifique contre l'aflatoxine. Le traceur peut se lier à l'anticorps monoclonal pour produire un changement détectable de la polarisation de fluorescence. Pour préparer le traceur on conjugue une oxime d'aflatoxine et un fluorophore approprié. On mesure la polarisation de fluorescence du mélange. On peut calculer la concentration des aflatoxines dans le mélange, à l'aide d'une courbe d'étalonnage obtenue par la mesure de la polarisation de fluorescence d'une série de solutions d'aflatoxines ayant une concentration connue.

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