Method for detecting analytes

• US 20060223060A1
• Filed: 09/30/2003
• Published: 10/05/2006
• Est. Priority Date: 09/30/2002
• Status: Abandoned Application

• Current Assignee: Cibitest GmbH & Company KG
• Original Assignee: Cibitest GmbH & Company KG
• Inventors: Hagenmaier, Sigrid, Polackova, Jitka, Kampen, Jan Von, Bonenberger, Johannes
• Application Number:
  US10/529,700
• Publication Number:
  US 20060223060A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

The present invention relates to a method and devices for detecting analytes comprising the following stages: incubation of a sample with macromolecules, to each of which at least 2 molecules of the analyte to be detected in the sample are coupled; subsequent incubation of the sample with a solid carrier, to which capture molecules for the analyte to be detected are coupled; addition of a fluorescence dye to stain the macromolecules and detection of analytes present in the sample by excitation of the fluorescence dye.

Claims

1. A method for detecting analytes comprising the following steps of:

a) incubating a sample with macromolecules, to each of which at least 2 molecules of the analyte to be detected in the sample are coupled;

b) further incubating the sample with a solid carrier, to which capture molecules for the analyte to be detected are coupled;

c) adding a fluorescence dye to stain the macromolecules; and

d) detecting the analytes present in the sample by excitation of the fluorescence dye.

2. The method according to claim 1 comprising, after step c), a further step c′

);

removing the non-bound fluorescence dye from the solid carrier.

5. The method according to claim 1, wherein the macromolecules are nucleic acids, peptide nucleic acids, polyamino acids.

6. The method according to claim 1, wherein the macromolecules are single-strand oligonucleotides of a length within the range from 40 to 80 nucleotides.

7. The method according to claim 1, wherein the macromolecules are identical or non-identical.

8. The method according to claim 1, wherein the analytes have a molecular weight of less than 5000 Dalton.

9. The method according to claim 1, wherein the fluorescence dye is selected from the group of phenanthrenes, acridines, SYBR dyes or fluorophores.

10. The method according to claim 1, wherein the solid carrier is permeable to light and the detection method is implemented by means of a transmitted-light method.

3. A method for detecting analytes comprising the steps:

a) incubating a sample with fluorescence-dye-marked macromolecules, to each of which at least 2 molecules of the analyte to be detected in the sample are coupled;

b) further incubating the sample with a solid carrier, to which capture molecules for the analyte to be detected are coupled; and

c) detecting analytes present in the sample by excitation of the fluorescence dye.

4. The method according to claim 3 comprising, after step a), a further step a′

);

removing the non-bound macromolecules.

12. The method according to claim 3, wherein the macromolecules are nucleic acids, peptide nucleic acids, polyamino acids.

13. The method according to claim 3, wherein the macromolecules are single-strand oligonucleotides of a length within the range from 40 to 80 nucleotides.

14. The method according to claim 3, wherein the macromolecules are identical or non-identical.

15. The method according to claim 3, wherein the analytes have a molecular weight of less than 5000 Dalton.

16. The method according to claim 3, wherein the fluorescence dye is selected from the group of phenanthrenes, acridines, SYBR dyes or fluorophores.

17. The method according to claim 3, wherein the solid carrier is permeable to light and the detection method is implemented by means of a transmitted-light method.

11. A device comprising a light source fitted on one side of a solid carrier inserted into the device, that a filter disposed respectively between the light source and the solid carrier on the other side of the solid carrier, wherein the device is designed in such a manner that light passing through the solid carrier passes through an aperture into the human eye or into an optical instrument.