ASSAY IMPLEMENTATION IN A MICROFLUIDIC FORMAT

• US 20070190525A1
• Filed: 12/29/2006
• Published: 08/16/2007
• Est. Priority Date: 06/02/2000
• Status: Active Grant

• Current Assignee: Honeywell International Inc.
• Original Assignee: Honeywell International Inc.
• Inventors: Padmanabhan, Aravind, Gu, Yuandong
• Granted Patent: US 7,553,453 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/143 Quality control, feedback systems
• CPC Class Codes: B01L 2200/146 Employing pressure sensors
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502776 specially adapted for focusing or laminating flows
• CPC Class Codes: B01L 9/527 for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/1484 microstructural devices
• CPC Class Codes: G01N 2015/0073 Red blood cells
• CPC Class Codes: G01N 2015/008 White cells
• CPC Class Codes: G01N 2015/0084 Platelets
• CPC Class Codes: G01N 2015/0088 Biological contaminants; Fouling
• CPC Class Codes: G01N 2015/012 : Red blood cells
• CPC Class Codes: G01N 2015/016 : White blood cells
• CPC Class Codes: G01N 2015/018 : Platelets
• CPC Class Codes: G01N 2015/019 : Biological contaminants; Fouling
• CPC Class Codes: G01N 2015/1486 : Counting the particles
• CPC Class Codes: G01N 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2333/445 : Plasmodium
• CPC Class Codes: G01N 2333/70514 : CD4
• CPC Class Codes: G01N 2333/70589 : CD45
• CPC Class Codes: G01N 2800/26 : Infectious diseases, e.g. generalised sepsis
• CPC Class Codes: G01N 33/5094 : for blood cell populations red blood cells G01N33/80
• CPC Class Codes: Y02A 50/30 : Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

An assay implementation in a microfluidic format in a cartridge relating to a point-of-care instrument platform for monitoring and diagnosing infectious diseases (e.g., AIDS and malaria). The platform may also provide a complete blood count. The instrument platform may hold the cartridge and a portion of an optical system for fluorescent and scattered light related analyses of blood sample in a flow channel of the cartridge.

Claims

1. A point of care analyzer comprising:

a portable instrument; and

a microfluidic cartridge insertable in the instrument; and

wherein;

the instrument and cartridge comprise;

a flow channel for conveying a blood sample;

at least one fluorescent optical channel for detecting and identifying markers on constituents of the blood sample for an immunoassay; and

at least one scattering optical channel for determining a count and/or properties of constituents of the blood sample; and

the constituents of the blood sample to be detected in an immunoassay may be at least one of a group of CD4, CD45, CDX, CDXX, p. falciparum, p. vivax, various other pathogens, and the like.

2. The analyzer of claim 1, wherein the scattering optical channel is for detecting light scattered by constituents of the sample at a large angle.

3. The analyzer of claim 2, wherein the instrument and cartridge further comprise:

a second scattering optical channel for detecting light scattered by constituents of the sample at a small angle; and

a third scattering optical channel for detecting light scattered by constituents of the sample at a forward angle.

4. The analyzer of claim 3, wherein light scattered by the constituents is for the determining a count and/or properties of constituents of the blood sample.

5. The analyzer of claim 1, wherein the cartridge further comprises:

a plurality of flow sensors; and

a plurality of reagent reservoirs.

6. A method for attaining a microfluidic-based assay on a cartridge, comprising:

providing a sample of whole blood to a microfluidic cartridge; and

performing a microfluidic-based assay on the cartridge situated in a portable instrument of an analyzer.

7. The method of claim 6, wherein performing the microfluidic-based assay comprises:

providing a whole-blood sample to an input of the cartridge;

lysing at least a portion of the sample;

establishing a supply of monoclonal antibodies in the cartridge for usage; and

doing at least one incubation step;

8. The method of claim 7, wherein a quantity of the sample is less than 25 micro liters.

9. The method of claim 7, wherein performing the microfluidic-based assay further comprises less than one dilution step.

10. The method of claim 7, wherein the supply of monoclonal antibodies is less than two micro liters.

11. The method of claim 7, wherein the assay uses less than one milliliter of lysing solution.

12. The method of claim 7, wherein the assay comprises less than three incubation steps.

13. The method of claim 12, wherein an incubation step occurs for a duration of less than one minute at about room temperature.

14. The method of claim 6, wherein a duration of the microfluidic-based assay is less than five minutes.

15. A point of care analyzer comprising:

an immunoassay module; and

a hematology module; and

wherein at least portions of the modules are situated in a cartridge insertable in an instrument of the analyzer.

16. The analyzer of claim 15, wherein:

the immunoassay module comprises a first microfluidic circuit;

the hematology module comprises a second microfluidic circuit; and

the first and second microfluidic circuits are situated in the cartridge.

17. The analyzer of claim 15, wherein:

the immunoassay module comprises a flow channel and at least one fluorescent optical channel proximate to the flow channel for determining and/or deducing a presence of CD4, CD8, CD45, CDX, CDXX, P. falciparum, P. vivax, TB, various other pathogens, and/or the like; and

the hematology module comprises a flow channel and at least one scattering optical channel proximate to the flow channel for determining at least a complete blood count.

18. The analyzer of claim 17, wherein the immunoassay module and the hematology module comprise the same flow channel.

19. The analyzer of claim 17, wherein the pathogens can effect AIDS, malaria, HIV, TB, and the like

20. The analyzer of claim 15, wherein the immunoassay module can label lymphocytes and monocytes with fluorescent markers or tags.

21. The analyzer of claim 17, wherein the hematology module can differentiate white blood cells into at least three categories.

22. The analyzer of claim 21, wherein the categories comprise lymphocytes, monocytes, neutrophils, eosinophils and basophils.

23. The analyzer of claim 17, wherein the hematology module and the immunoassay module utilize a single common light source.

24. The analyzer of claim 16, wherein the microfluidic circuits comprise:

a hydrodynamic focusing channel;

a waste reservoir;

reagent reservoirs (sheathing, lysing, diluent); and

an anti-body reservoir.

25. A method for blood analysis comprising:

performing a hematology test; and

performing a immunoassay; and

wherein the test and assay are performed with a single microfluidic cartridge.

26. A method of claim 25, wherein the hematology test comprises:

providing a sample of whole blood; and

performing a complete blood count; and

wherein the complete blood count comprises;

lysing a first portion of the sample to result in white blood cells;

hydrodynamically focusing and pushing a first portion of the white blood cells through a channel to scatter light;

differentiating the white blood cells into types according to light scattered by the white blood cells;

hydrodynamically focusing a second portion of the sample through a channel to scatter light;

counting the red blood cells according to light scattered by the red blood cells;

determining hematocrit data from light scattered by the red blood cells; and

determining hemoglobin content of a third portion of the sample with light absorption.

27. The method of claim 26, wherein the immunoassay comprises:

incubating and tagging a second portion of the white blood cells with fluorescent antibodies;

impinging the white blood cells with light;

detecting fluorescence emitted from the tagged white blood cells;

detecting light scattered by the white blood cells; and

determining CDXX and/or pathogen data.

28. The method of claim 27, wherein the lysing, differentiation, hydrodynamically focusing, tagging and counting cells are performable on the single microfluidic cartridge.

29. The method of claim 28, wherein the incubating and determining CDXX and/or pathogen data are performable on the single microfluidic cartridge.

30. An assay system comprising:

a card; and

an immunoassay component situated in the card.

31. The system of claim 30, further comprising a hematology test component situated in the card.

32. The system of claim 31, wherein the card is a microfluidic cartridge comprising plastic type and/or glass type materials.

33. The system of claim 32, wherein the card is a laminated structure.

34. The system of claim 32, wherein the card has a molded structure.

35. The system of claim 31, wherein:

the immunoassay component can perform a CDXX and/or pathogen assay indicative of AIDS and/or malaria, respectively; and

the hematology test component can perform a blood count.

36. The system of claim 31 wherein the card comprises microfluidic circuits for mixing, separation, hydrodynamic focusing, filtering, wash-out, preconcentration, lysing, sorting, counting, incubating, tagging, and/or the like, for immunoassays and hematology tests.