Patent ID: 7476549
Filing Date: 2009-01-13
Classification: B01L,G01N,G02B

Abstract:
1. An lateral flow quantitative assay method in which a liquid sample which is expected to contain an analyte is applied at one end of a chromatography medium to move through the chromatography medium, such that the analyte in the sample reacts with a labeled detector adsorbed on a section located at a predetermined distance from the sample application in the sample developing direction, thereby forming an analyte/labeled detector conjugate; in which the analyte/labeled detector conjugate, while moving through the chromatography medium, further reacts with an unlabeled captor which is different from or identical to the detector and is immobilized on a viewing window defined around middle portion of the chromatography medium, thereby forming a labeled detector/analyte/unlabeled captor conjugate in a sandwich configuration; and in which an amount of the conjugate is measured for quantitative determination of the analyte in the sample, characterized in that: (a) the labeled detector is labeled with fluorescent material and reacts with the analytes in the liquid sample to form a fluorescently-labeled detector/analyte conjugate; (b) the unlabeled captor is immobilized in lines within the viewing window on the chromatography medium and reacts with the fluorescently-labeled detector/analyte conjugate which has moved along the chromatography medium to form a fluorescently-labeled detector/analyte/unlabeled captor conjugate; (c) a reference detector which is different from the detector and captor, is labeled with the same fluorescent material as the detector and reacts with reference material in the liquid sample, is adsorbed on the section of the chromatography medium where the fluorescently-labeled detector is adsorbed, and an unlabeled reference captor which reacts with the fluorescently-labeled reference detector is immobilized in a single reference line before or after the viewing window on the chromatography medium, or in double reference lines before and after the viewing window on the chromatography medium, whereby a reference conjugate of fluorescently-labeled reference detector/reference material/unlabeled reference captor is formed as the liquid sample passes through the chromatography medium; (d) said chromatography medium is comprised of two layers of laminated chromatography media, and said captor and the reference captor are immobilized on the chromatography medium and/or between said two layers of laminated chromatography media; and (e) an amount of the analyte is determined by passing light emitted from a laser through an exciter filter, irradiating the filtered light to the epifluorescence medium containing the analyte conjugate and the reference conjugate, focusing light reflected from epifluorescence medium to a first focal point of an elliptical or spherical reflecting mirror with a proper size, focusing scattered incident light and fluorescence emitted from the sample positioned at the first focal point of the elliptical reflecting mirror to a second focal point of the elliptical reflecting mirror, converting the focused light into parallel light by a collimator, filtering the parallel light through a fluorescent filter to remove the scattered incident light and provide a pure a fluorescence component to an optical detector, and comparing a fluorescence intensity of the analyte conjugate with a reference fluorescence intensity of the reference conjugate to determine the relative amount of the analyte.