ULTRA-SENSITIVE DETECTION OF MOLECULES OR PARTICLES USING BEADS OR OTHER CAPTURE OBJECTS

• US 20120277114A1
• Filed: 06/22/2012
• Published: 11/01/2012
• Est. Priority Date: 03/01/2010
• Status: Active Grant

• Current Assignee: Quanterix Corporation
• Original Assignee: Quanterix Corporation
• Inventors: Duffy, David C., Rissin, David M., Walt, David R., Fournier, David, Kan, Cheuk
• Granted Patent: US 9,310,360 B2
• Time in Patent Office: Days
• US Class Current: 506/9
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• 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

Abstract

The present invention relates to systems and methods for detecting analyte molecules or particles in a fluid sample and in some cases, determining a measure of the concentration of the molecules or particles in the fluid sample. Methods of the present invention may comprise immobilizing a plurality of analyte molecules or particles with respect to a plurality of capture objects. At least a portion of the plurality of capture objects may be spatially separated into a plurality of locations. A measure of the concentration of analyte molecules in a fluid sample may be determined, at least in part, on the number of reaction vessels comprising an analyte molecule immobilized with respect to a capture object. In some cases, the assay may additionally comprise steps including binding ligands, precursor labeling agents, and/or enzymatic components.

Claims

1. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising:

providing a substrate comprising a plurality of locations, at least a portion of which locations contain a bead, wherein with respect to the total number of beads present on the substrate, the ratio of beads comprising at least one analyte molecule or particle to beads comprising no analyte molecules or particles is between about 8;

1 and about 1;

10,000,000;

addressing at least a portion of the plurality of locations, wherein during the addressing step at least two of the plurality of locations is addressed at least partially concurrently;

detecting at each addressed location the presence or absence of a bead and whether, if present, the bead comprises any analyte molecules or particles; and

determining a measure of the concentration of analyte molecules or particles in the fluid sample at least in part by determining the number of locations addressed containing a bead comprising at least one analyte molecule or particle.

2. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising:

providing a substrate comprising a plurality of locations, at least a portion of which contain a bead, wherein with respect to the total number of beads present on the substrate, the ratio of beads comprising at least one analyte molecule or particle associated with a binding ligand to beads comprising no analyte molecules or particles associated with a binding ligand is between about 8;

1 and about 1;

10,000,000;

addressing at least a portion of the plurality of locations, wherein during the addressing step at least two of the plurality of locations is addressed at least partially concurrently;

detecting at each addressed location the presence or absence of a bead and whether, if present, the bead comprises any analyte molecules or particles associated with a binding ligand; and

determining a measure of the concentration of analyte molecules or particles in the fluid sample at least in part by determining the number of locations addressed containing a bead comprising at least one analyte molecule or particle associated with a binding ligand.

3. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising:

immobilizing a plurality of analyte molecules or particles with respect to a plurality of beads;

spatially segregating at least a portion of the plurality of beads into a plurality of separate locations;

addressing at least some of the plurality of locations and determining the number of locations containing a bead;

further determining the number of said locations containing a bead and an analyte molecule or particle; and

determining a measure of the concentration of analyte molecules or particles in the fluid sample based at least in part on the ratio of the number of locations containing a bead and an analyte molecule and particle, to the number of locations containing a bead.

4. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising:

immobilizing a plurality of analyte molecules or particles with respect to a plurality of beads;

spatially segregating at least a portion of the plurality of beads into a plurality of separate locations;

addressing at least some of the plurality of locations and determining the number of locations containing a bead;

further determining the number of said locations containing a bead and an analyte molecule or particle; and

determining a measure of the concentration of analyte molecules or particles in the fluid sample based at least in part on the ratio of the number of locations containing a bead and an analyte molecule and particle, to the number of locations containing a bead but not containing any analyte molecules or particles.