Particle on membrane assay system

• US 20050191620A1
• Filed: 12/22/2004
• Published: 09/01/2005
• Est. Priority Date: 02/27/2004
• Status: Active Grant

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Ballard, Karri L., McDevitt, John T., Bernard, Bruce, Floriano, Pierre, Christodoulides, Nick
• Granted Patent: US 7,781,226 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: G01N 2021/0321 One time use cells, e.g. integrally moulded
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 33/521 Single-layer analytical elements
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/805 Optical property

Abstract

Described herein is an analyte detection device and method related to a portable instrument suitable for point-of-care analyses. In some embodiments, a portable instrument may include a disposable cartridge, an optical detector, a sample collection device and/or sample reservoir, reagent delivery systems, fluid delivery systems, one or more channels, and/or waste reservoirs. Use of a portable instrument may reduce the hazard to an operator by reducing an operator'"'"'s contact with a sample for analysis. The device is capable of obtaining diagnostic information using cellular- and/or particle-based analyses and may be used in conjunction with membrane- and/or particle-based analysis cartridges. Analytes, including proteins and cells and/or microbes may be detected using the membrane and/or particle based analysis system.

Claims

1. A method of detecting an analyte in a fluid comprising:

forming a mixture of one or more particles with the fluid, wherein each particle comprises a receptor configured to interact with the analyte, and wherein particles that interact with the analyte undergo a detectable spectroscopic change;

passing the mixture of particles and fluid across a porous membrane disposed in an analyte detection device, wherein the analyte detection device is configured to capture the particles on the porous membrane, wherein the analyte detection device comprises;

a porous membrane;

a top member positioned at a spaced distance above the porous membrane such that a first cavity is formed between the top member and the porous membrane, wherein the top member covers at least a portion of the porous membrane, and wherein the top member is substantially transparent to light, and

a bottom member positioned below the porous membrane, wherein the bottom member is configured to receive fluid flowing through the porous membrane during use;

detecting spectroscopic signals produced by particles captured on the porous membrane; and

determining if the analyte is present based on the detected spectroscopic signals.

2. The method of claim 1, wherein each of the particles comprises an identification molecule, wherein the identification molecule coupled to each particle is specific for the analyte that interacts with the particle, the method further comprising determining if the analyte is present by determining the identity of the identification molecule on particles in which a spectroscopic signal has been detected.

3. The method of claim 1, wherein each of the particles comprises a predetermined size, wherein the size of each particle is specific for the analyte that interacts with the particle, the method further comprising determining if the analyte is present by determining the size of the particles in which a spectroscopic signal has been detected.

5. The method of claim 1, further comprising detecting additional analytes, wherein each analyte is detected by one or more particles that are configured to produce a spectroscopic signal when one or more particles interact with the additional analytes.

6. The method of claim 1, wherein forming a mixture of one or more particles with the fluid further comprises adding a detecting receptor to the fluid, wherein the particles are configured to produce a first signal, and wherein the detecting receptor is configured to produce a second signal, and wherein determining if the analyte is present comprises detecting the first and second signals.

21. The method of claim 1, wherein the top member comprises a fluid inlet configured to allow fluid to be introduced to the membrane through the top member.

22. The method of claim 1, wherein the bottom member comprises a fluid outlet configured to allow fluid to pass from the membrane out of the analyte detection device.

28. The method of claim 1, wherein the receptor comprises an antibody that binds to at least a portion of a virus.

30. The method of claim 1, wherein the receptor comprises an antibody that binds to at least a portion of the human immunodeficiency virus.

36. The method of claim 1, wherein the receptor comprises an antibody to the analyte.

37. The method of claim 1, further comprising adding a visualization agent to the particles captured on the porous membrane.

39. The method of claim 1, further comprising passing a background fluid through the analyte detection device and detecting an image captured on the porous membrane prior to passing the fluid containing the analyte across the porous membrane.

40. The method of claim 1, further comprising washing the surface of the porous membrane.

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