Methods for multiplex analyte detection and quantification

G - Physics – 01 – N

Patent

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Details

G01N 33/53 (2006.01) C40B 30/04 (2006.01) G01N 33/543 (2006.01) G01N 33/564 (2006.01) G01N 33/58 (2006.01)

Patent

CA 2748707

The application refers to a method for detecting and quantifying multiple target ana-lytes in a test sample using a single reaction vessel. The method uses a reaction vessel (a multi-well plate), which comprises a microarray of : (a) cali-bration spots, each having a predetermined quanti-ty of the target analyte; and (b) capture spots, each having an agent (antibody) that selectively binds the target analyte. The captured analytes and the calibration spots are detected with fluorescently la-belled antibodies specific for each different target analyte. The calibration spots are used to generate calibration curves that allow the measurement of the concentration of the different target analytes. The application also refers to a method for detect-ing and quantifying biomarkers that are useful for diagnosing rheumatoid arthritis. More specifically, the application discloses the use of rheumatoid fac-tor (RF) and cyclic citrullinated peptide (CCP), as capture spots. Finally, based on the above method, it is proposed a method for diagnosing or monitor-ing rheumatoid arthritis.

L'invention porte sur un procédé de détection et de quantification d'analytes cibles multiples dans un échantillon test à l'aide d'un seul récipient de réaction. Le procédé utilise un récipient de réaction (une plaque à puits multiples), qui comprend un microréseau de : (a) points d'étalonnage, chacun ayant une quantité prédéterminée de l'analyte cible ; et (b) points de capture, ayant chacun un agent (anticorps) qui se lie sélectivement à l'analyte cible. Les analytes capturés et les points d'étalonnage sont détectés par des anticorps à étiquetage fluorescent spécifiques pour chaque analyte cible différent. On utilise les points d'étalonnage pour générer des courbes d'étalonnage permettant la mesure de la concentration des différents analytes cibles. L'invention porte également sur un procédé de détection et de quantification de biomarqueurs qui sont utiles pour le diagnostic de la polyarthrite rhumatoïde. Plus spécifiquement, l'invention porte sur l'utilisation d'un facteur rhumatoïde (RF) et d'un peptide citrulliné cyclique (CCP) en tant que points de capture. Enfin, sur la base du procédé ci-dessus, on propose un procédé de diagnostic ou de surveillance de la polyarthrite rhumatoïde.

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