Lateral flow test format for enzyme assays

• US 20040219694A1
• Filed: 03/10/2004
• Published: 11/04/2004
• Est. Priority Date: 09/11/2001
• Status: Abandoned Application

• Current Assignee: Merck Patent GmbH (Merck & Co., Inc.)
• Original Assignee: Merck Patent GmbH (Merck & Co., Inc.)
• Inventors: Watkins, Ian David, Chittock, Roger Steward
• Application Number:
  US10/489,166
• Publication Number:
  US 20040219694A1
• Time in Patent Office: Days
• US Class Current: 436/518
• CPC Class Codes: C12Q 1/008 for determining co-enzymes or co-factors, e.g. NAD, ATP
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387

Abstract

The invention relates to a lateral flow enzyme assay device and test kit for the determination of analyte in a test sample. The invention further relates to lateral flow method for the determination of analytes by directly using the analyte as an enzyme substrate.

Claims

1. Method for determining the presence of an analyte in a sample comprising:

a) providing a test device incorporating a lateral flow test strip comprising a dry matrix material capable of transporting a liquid therealong by capillarity and having at least a start zone for receiving said sample and a reaction zone having at least one kind of enzyme immobilised therein;

b) contacting the start zone with said sample thereby causing a liquid phase to move through said test strip;

c) determining a detectable signal at the reaction zone directly or indirectly caused by covalent modification of the analyte by the enzymes at the reaction zone

2. Method according to claim 1, characterised in that the test strip provided in step a) comprises an additional reagent zone.

3. Method according to claim 1 characterised in that the test strip provided in step a) comprises additional dried reagents in the start zone and/or the reagent zone and/or the reaction zone.

4. Method according to claim 1, characterised in that the liquid flow generated in step b) is enhanced by a wick on the test strip.

5. Method according to claim 1, characterised in that the detectable signal is generated by a product of an enzymatic reaction which is visible by eye and barely soluble.

6. Method according to claim 1, characterised in that the detectable signal in step c) is generated by using a recycling enzyme reaction.

7. Lateral flow test device for determining the presence of an analyte in a sample incorporating a test strip comprising a dry matrix material capable of transporting a liquid therealong by capillarity and having at least a start zone for receiving said sample and a reaction zone having at least one kind of enzyme immobilised therein whereby the enzymes and optional additional reagents on the test strip are suitable for covalently modifying the analyte and thus directly or indirectly generating a detectable signal.

8. Lateral flow test device according to claim 7, characterised in that the test strip further comprises a solid support on which is attached the dry matrix material.

9. Lateral flow test device according to claim 7, characterised in that the test strip further comprises a reagent zone containing dried reagents.

10. Lateral flow test device according to claim 7, characterised in that the test strip comprises a start zone containing a dried detergent extractant, a reagent zone containing one or more dried enzyme substrates and/or cofactors and a reaction zone containing at least one kind of immobilised enzyme.

11. Lateral flow test device according to claim 7, characterised in that the test strip further comprises a wick.

12. Lateral flow test device according to claim 7, characterised in that it further comprises a housing.

13. Lateral flow test device according to claim 7, characterised in that the reaction and/or reagent zones are protected by a transparent film to allow ease of handling and prevent contamination while allowing viewing of the signal formed at the reaction zone

14. Lateral flow test device according to claim 7, characterised in that the test strip comprises a start zone containing DTAB (Dodecyl Trimethyl Ammonium Bromide) as detergent extractant, a reagent zone containing the substrates NBT and glucose and a reaction zone containing the immobilised enzymes glucose dehydrogenase and diaphorase.

15. Use of a lateral flow test device according to claim 7 in an enzyme assay.

16. Use of a lateral flow test device according to claim 14 to test surfaces.

17. Use of a lateral flow test device according to claim 14 to test liquids.

18. Enzyme assay test kit comprising a lateral flow test device according to claim 7.