Moving droplet diagnostic assay

• US 8,586,349 B2
• Filed: 08/06/2001
• Issued: 11/19/2013
• Est. Priority Date: 08/10/2000
• Status: Expired due to Term

• Current Assignee: Biomerieux B.V. (Compagnie Merieux Alliance SAS)
• Original Assignee: Biomerieux B.V. (Compagnie Merieux Alliance SAS)
• Inventors: Beumer, Thomas Augustinus Maria, Verheijden, Petrus Franciscus Hendrikus M., van Damme, Hendrik Sibolt
• Primary Examiner(s): Hines, J.
• Application Number:
  US10/344,226
• Publication Number:
  US 20040043421A1
• Time in Patent Office: 4,488 Days
• Field of Search: 422/55, 422/56, 422/57, 422/58, 422/61, 422/68.1, 435/7.2, 435/283.1, 435/286.5, 435/287.1, 435/287.2, 435/287.9, 435/288.5, 435/288.7, 436/517, 436/518, 436/523, 436/536
• US Class Current: 435/287.1
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: Y10T 436/2525 Stabilizing or preserving

Abstract

The present invention concerns a method for the detection of an analyte in a droplet of fluid comprising the steps of providing a solid phase with a hydrophobic surface comprising at least one capture zone on which at least one binding agent with affinity for the analyte is immobilized, applying said droplet to the surface of said solid phase, applying a force that makes the droplet travel along the surface of said solid phase along a predetermined path thereby allowing the droplet to repeatedly contact said binding agent on the capture zone, applying conditions wherein said analyte is allowed to bind to said binding agent and detecting a complex of analyte and binding reagent at the position of the capture zone. The effect of a moving droplet is that the reactants in the droplet are well mixed which eliminates the risk of diffusion limitation. Such advantageous mixing is not obtained with conventional assays in which the surface is continuously exposed to the liquid. The use of a droplet eliminates the need for external mixing and also circumvents the need to dilute the sample in order to obtain sufficient volume to wet a large surface area.

Claims

1. A device for the detection of an analyte in a droplet of fluid, comprising:

a non-porous and hydrophobic solid phase surface configured to receive and maintain the shape of a droplet of fluid;

a circumferential gutter in said solid phase surface that defines a predetermined continuous pathway; and

a capture zone located in the gutter;

wherein said capture zone includes an immobilized binding agent positioned in said capture zone, said gutter configured such that when said droplet is positioned in said gutter and traveling along said pathway, said droplet repeatedly and periodically travels into and out of said capture zone.

2. The device of claim 1, wherein said gutter is provided with at least one obstacle.

3. The device of claim 1, wherein said gutter is circular.

4. The device of claim 1, wherein said circumferential gutter comprises a plurality of capture zones located in the gutter with each capture zone including an immobilized binding agent therein, the gutter being configured such that said droplet repeatedly and periodically travels into and out of said capture zone.

5. A kit comprising:

a) the device of claim 1;

b) reagents for detecting an analyte in the droplet of fluid; and

c) instructions for use.

6. The kit of claim 5, wherein said gutter is provided with at least one obstacle.

7. A kit comprising:

a) the device of claim 3;

b) reagents for detecting an analyte in the droplet of fluid; and

c) instructions for use.

8. A method for the detection of an analyte in a droplet of fluid, comprising the steps of:

a) providing a non-porous and hydrophobic solid phase surface for receiving and maintaining the shape of said droplet, said surface comprising a circumferential gutter defining a predetermined continuous pathway and wherein a capture zone that includes an immobilized binding agent is positioned in said gutter;

b) applying said droplet to said gutter;

c) applying a force that makes said droplet travel along said predetermined continuous pathway of said gutter, wherein said droplet repeatedly and periodically travels into and out of said capture zone while maintaining a droplet shape, resulting in the formation of a complex of said analyte with said immobilized binding agent, further wherein the amount of complex increases as said droplet repeatedly and periodically travels into and out of said capture zone; and

d) detecting said complex of analyte and binding agent in said capture zone, thereby detecting said analyte in said droplet of fluid.

9. The method of claim 8, wherein manual force is used to make the droplet travel along said predetermined continuous pathway of said gutter.

10. The method of claim 8, wherein the analyte is selected from the group consisting of antigens, antibodies, haptens, nucleic acids, cells, membranes, bacteria and viruses.

11. The method of claim 8, wherein the binding agent is selected from the group consisting of antigens, antibodies, peptides, nucleic acids and polymers.

12. The method of claim 8, wherein said gutter is provided with at least one obstacle.

13. The method of claim 8, wherein said gutter is circular.

14. The method of claim 8, wherein the force applied to make the droplet travel along said predetermined continuous pathway of said gutter is a gravitational force.

15. The device of claim 14, wherein the plurality of said capture zones comprise the same binding agent.

16. The device of claim 14, wherein the plurality of said capture zones comprise different binding agents.