Method and test kit for detecting analytes in a sample

• US 20040106215A1
• Filed: 12/09/2003
• Published: 06/03/2004
• Est. Priority Date: 10/27/2000
• Status: Active Grant

• Current Assignee: Attomol GmbH Molekulare Diagnostika
• Original Assignee: Attomol GmbH Molekulare Diagnostika
• Inventors: Lehmann, Werner
• Granted Patent: US 7,361,515 B2
• Time in Patent Office: Days
• US Class Current: 436/518
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention relates to a method and test kit for the detection of analytes in a biological sample, wherein at least one microparticle population is labelled with fluorescent dyes serving as coding fluorescent dyes and reference fluorescent dyes and immobilized, acceptor molecules interacting with labelled analytes being bound to the microparticle population, so that the difference between the fluorescence of the microparticles and analyte(s) can be used to determine which kind of analyte, optionally in which amount, is present in the sample. In particular, the invention finds use in medical diagnostics.

Claims

1. A method for the detection of analytes in a sample, characterized in that the method comprises the following steps:

fluorescent labelling of at least one microparticle population, said microparticle population being labelled with at least one fluorescent dye serving as coding fluorescence and at least one fluorescent dye serving as reference fluorescence,

binding and/or conjugating acceptor molecules to the microparticle population,

immobilizing the microparticle population on a support,

incubating the microparticle population with the sample,

labelling the analyte(s) with at least one reporter fluorescence, and

detecting the analyte(s) by comparing the fluorescence of the microparticles with the reporter fluorescence.

2. The method according to claim 1,

characterized in that the microparticles are immobilized on microtest plates, glass slides, flexible membranes, networks, fibrils, particularly those made of polypropylene and/or nitrocellulose, glass and/or PVDF.

3. The method according to any of the preceding claims, characterized in that a plurality of analytes are detected simultaneously by loading each microparticle population with individual acceptor molecules.

4. The method according to any of the preceding claims, characterized in that the analytes are labelled with different or identical reporter fluorescences.

5. A test kit,

characterized in that the test kit comprises at least one immobilized microparticle population conjugated with specific acceptor molecules, whereby the microparticle population comprises at least one coding fluorescence and at least one reference fluorescence differing in their spectral properties and/or their lifetime of fluorescence.

6. The test kit according to the preceding claim, characterized in that the microparticle population comprises at least one coding fluorescence and at least one reference fluorescence.

7. Use of at least one immobilized, fluorescent-labelled microparticle population having specifically conjugated acceptor molecules, said microparticle population comprising a coding fluorescence and a reference fluorescence, in the detection of analytes in clinical and/or environmental diagnostics.