Method and system for the analysis of saliva using a sensor array

• US 7,651,868 B2
• Filed: 12/13/2004
• Issued: 01/26/2010
• Est. Priority Date: 12/11/2003
• 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: Christodoulides, Nick J., Neikirk, Dean P., Anslyn, Eric V., McDevitt, John T., Shear, Jason B.
• Primary Examiner(s): Shibuya; Mark L
• Assistant Examiner(s): Diramio; Jacqueline
• Application Number:
  US11/010,816
• Publication Number:
  US 20050214863A1
• Time in Patent Office: 1,870 Days
• Field of Search: 422/55, 422/58, 422/68.1, 422/82.05, 422/82.08, 4352871-2872, 4352883-2884, 435/288.5, 435/288.7, 436/501, 436/518, 436/523, 436/531, 436/532, 436/164, 436/172, 436/71, 436/85, 436/807, 436/809
• US Class Current: 436/518
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 2021/7783 : Transmission, loss
• CPC Class Codes: G01N 2021/7786 : Fluorescence
• CPC Class Codes: G01N 21/6428 : Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6454 : using an integrated detector array
• CPC Class Codes: G01N 21/77 : by observing the effect on a chemical indicator
• CPC Class Codes: G01N 2201/024 : Modular construction
• CPC Class Codes: G01N 2800/32 : Cardiovascular disorders
• CPC Class Codes: G01N 33/5304 : Reaction vessels, e.g. agglutination plates for solid-phase systems G01N33/543
• CPC Class Codes: G01N 33/54313 : the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/544 : the carrier being organic
• CPC Class Codes: G01N 33/552 : Glass or silica
• CPC Class Codes: G01N 33/6887 : from muscle, cartilage or connective tissue
• CPC Class Codes: G01N 33/92 : involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743
• CPC Class Codes: Y10S 436/807 : Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 : Multifield plates or multicontainer arrays

Abstract

A system for the rapid characterization of analytes in saliva. In one embodiment, a system for detecting analytes includes a light source, a sensor array, and a detector. The sensor array is formed from a supporting member, in which a plurality of cavities may be formed. A series of chemically sensitive particles, in one embodiment, are positioned within the cavities. The particles may produce a signal when a receptor, coupled to the particle, interacts with the cardiovascular risk factor analyte and the particle-analyte complex is visualized using a visualization reagent. Using pattern recognition techniques, the analytes within a multi-analyte fluid may be characterized. In an embodiment, each cavity of the plurality of cavities is designed to capture and contain a specific size particle. Flexible projections may be positioned over each of the cavities to provide retention of the particles in the cavities.

Claims

1. A method for detecting one or more cardiovascular risk factor analytes in saliva comprising:

passing saliva over a sensor array, the sensor array comprising;

a supporting member comprising a plurality of cavities formed within the supporting member;

a plurality of particles, at least one of the particles positioned in at least one of the cavities, wherein at least one of the particles is configured to produce a signal in the presence of at least one of the cardiovascular risk factor analytes during use, wherein at least one of the particles is configured to (a) produce a signal in the presence of C-reactive protein and (b) exhibit a sensitivity sufficient for detecting C-reactive protein levels in saliva of less than 1.9 ng/ml; and

monitoring a spectroscopic change of at least one of the particles as the saliva is passed over the sensor array.

2. The method of claim 1, wherein the sensor array further comprises a bottom layer and a cover, wherein the bottom layer is coupled to a bottom surface of the supporting member, and wherein the cover is coupled to a top surface of the supporting member;

and wherein both the bottom layer and the cover are coupled to the supporting member such that at least one of the particles is substantially contained in at least one of the cavities by the bottom layer and the cover, and wherein the bottom layer and the cover are substantially transparent to light produced by a light source.

3. The method of claim 1, wherein the sensor array further comprises a cover, the cover being coupled to the supporting member such that at least one of the particles is substantially contained in at least one of the cavities by the cover, and wherein the cover is configured to allow the saliva to pass through the cover to at least one of the particles, and wherein the supporting member and the cover are substantially transparent to light produced by a light source.

4. The method of claim 1, wherein the sensor array further comprises a cover positioned at a distance above the upper surface of the supporting member such that an opening is formed between the supporting member and the cover to allow the saliva to enter at least one of the cavities via the opening, and wherein the cover inhibits dislodgment of at least one of the particles from at least one of the cavities during use.

5. The method of claim 1, wherein at least one of the cavities is configured such that the saliva entering the cavity passes through the supporting member during use.

6. The method of claim 1, wherein at least one of the cavities is substantially tapered such that the width of the cavity narrows in a direction from a top surface of the supporting member toward a bottom surface of the supporting member, and wherein a minimum width of the cavity is substantially less than a width of at least one of the particles.

7. The method of claim 1, wherein at least one of the particles comprises a receptor molecule coupled to a polymeric resin.

8. The method of claim 1, wherein at least one of the particles comprises a receptor molecule coupled to a polymeric resin, and wherein the receptor molecule comprises a peptide.

9. The method of claim 1, wherein at least one of the particles comprises a receptor molecule coupled to a polymeric resin, and wherein the receptor molecule comprises a synthetic receptor.

10. The method of claim 1, wherein at least one of the particles comprises a receptor molecule coupled to a polymeric resin, and wherein the receptor molecule comprises an antibody.

11. The method of claim 1, wherein at least a portion of the particle comprises a receptor molecule coupled to a polymeric resin, and wherein the receptor molecule comprises an antigen.

12. The method of claim 1, wherein at least one of the cardiovascular risk factor analytes is interleukin-6.

13. The method of claim 1, wherein at least one of the cardiovascular risk factor analytes is high density lipoprotein, low density lipoprotein, very low density lipoprotein, cholesterol, interleukin-6, intercellular adhesion molecule-1, fibrinogen, homocysteine, folate, calcium, lipoprotein a, apolipoprotein A-1, apolipoprotein B, Helicobacter pylon, chlamydia pneumoniae, Herpes virus hominis, or cytomegalovirus.

14. The method of claim 1, further comprising simultaneously detecting the presence of two or more cardiovascular risk factor analytes in saliva.

15. The method of claim 1, further comprising simultaneously detecting the presence of two or more cardiovascular risk factor analytes in saliva, wherein one of the cardiovascular risk factor analytes is high density lipoprotein, low density lipoprotein, very low density lipoprotein, cholesterol, interleukin-6, intercellular adhesion molecule-1, fibrinogen, homocysteine, folate, calcium, lipoprotein a, apolipoprotein A-1, apolipoprotein B, Helicobacter pylon, chlamydia pneumoniae, Herpes virus hominis, or cytomegalovirus.

16. The method of claim 1, wherein two or more of the particles are configured to produce a detectable signal in the presence of one or more cardiac risk factor analytes.

17. The method of claim 1, wherein the supporting member comprises silicon.

18. The method of claim 1, wherein the sensor array further comprises channels in the supporting member, wherein the channels are configured to allow the saliva to flow through the channels into and away from at least one of the cavities.

19. The method of claim 1, wherein the sensor array further comprises a pump coupled to the supporting member, wherein the pump is configured to direct the saliva towards at least one of the cavities, and wherein a channel is formed in the supporting member, the channel coupling the pump to at least one of the cavities such that the saliva flows through the channel to at least one of the cavities during use.

20. A method for detecting two or more cardiovascular risk factor analytes in saliva comprising:

passing saliva over a sensor array, the sensor array comprising;

a supporting member comprising two or more cavities formed within the supporting member; and

two or more particles, at least a first one of the particles positioned in a first one of the cavities and a second one of the particles positioned in a second one of the cavities, wherein at least the first one of the particles is configured to (a) produce a signal in the presence of C-reactive protein during use and (b) exhibit a sensitivity sufficient for detecting C-reactive protein levels in saliva of less than 1.9 ng/ml, and wherein at least the second one of the particles is configured to produce a signal in the presence of a cardiovascular risk factor analyte different than C-reactive protein during use; and

monitoring a spectroscopic change of at least one of the particles as the saliva is passed over the sensor array.