Microfluidic apparatus and methods for performing blood typing and crossmatching

• US 10,107,797 B2
• Filed: 08/19/2015
• Issued: 10/23/2018
• Est. Priority Date: 10/03/2008
• Status: Active Grant

• Current Assignee: Revvity Health Sciences Incorporated (Revvity, Inc.)
• Original Assignee: Micronics Inc. (Sony Group Corp.)
• Inventors: Battrell, C. Frederick, Wierzbicki, Diane, Clemmens, John, Capodanno, Jason, Williford, John R., Elmufdi, Carolina, Sprague, Isaac
• Primary Examiner(s): Martinez, Rebecca L
• Application Number:
  US14/830,539
• Publication Number:
  US 20160209431A1
• Time in Patent Office: 1,161 Days
• Field of Search: None
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• 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 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/161 Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• 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: B01L 3/502776 specially adapted for focusing or laminating flows
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/5304 Reaction vessels, e.g. agglutination plates for solid-phase systems G01N33/543
• CPC Class Codes: G01N 33/80 involving blood groups or blood types or red blood cells white blood cells G01N33/56972

Abstract

Microfluidic cartridges for agglutination reactions are provided. The cartridges include a microfluidic reaction channel with at least two intake channels, one for an antigen-containing fluid and the other for an antibody-containing fluid, conjoined to a reaction channel modified by incorporation of a downstream flow control channel. At low Reynolds Number, the two input streams layer one on top of the other in the reaction channel and form a flowing, unmixed horizontally-stratified laminar fluid diffusion (HLFD) interface for an extended duration of reaction. Surprisingly, the design, surface properties, and flow regime of microfluidic circuits of the present invention potentiate detection of antibody mediated agglutination at the stratified interface. Antigen:antibody reactions involving agglutination potentiated by these devices are useful in blood typing, in crossmatching for blood transfusion, and in immunodiagnostic agglutination assays, for example.

Claims

1. A microfluidic cartridge for performing a blood type assay or crossmatch, which comprises:

a) a body member comprised of a substrate or substrates, said body member enclosing a reaction channel, said reaction channel having a first end and a second end and having a critical dimension configured for establishing laminar flow conditions;

wherein said body member is comprised of an upper body layer and a base body layer, wherein said upper body layer and base body layer are joined by an adhesive core adhesive (ACA) glue layer, said ACA glue layer having a first glue face and a second glue face separated by a supportive core, and further wherein said first glue face and said second glue face of said ACA glue layer are formulated with an additive selected to modify the contact angle of the glue faces, wherein the contact angle of said first glue face is configured for wetting with whole blood and the contact angle of said second glue face is configured for wetting with a reagent solution, and thereby said contact angle of the first glue face and the contact angle of the second glue face are measurably distinct, wherein said contact angle of the first glue face is in the range of 10 to 20 degrees and said contact angle of the second glue face is in the range of 20 to 30 degrees;

b) a first and a second intake channel fluidly joined to said first end of said reaction channel at a staging union;

said first intake channel for conveying a first fluid and said second intake channel for conveying a second fluid;

wherein said staging union is configured with a dual capillary stop for simultaneously releasing said first and second fluids into said reaction channel in response to a start signal, said fluids contactedly flowing in parallel within said reaction channel as lamellae, one on top of the other;

c) a downstream flow control channel fluidly joined to the second end of said reaction channel, wherein the downstream flow control channel is further characterized as comprising an elongate runoff channel segment configured with microfluidic dimensions and reverse tapered walls that broaden from upstream to downstream for sustaining and modulating a passive laminar flow regime in said reaction channel wherein said passive laminar flow regime in said reaction channel is modulated and sustained as a horizontally-stratified laminar fluid diffusion interface characterized by an apparent Reynolds Number between 0.1 and 10, as determined in said reaction channel, for a duration of about thirty seconds to ten minutes, more preferably about thirty seconds to two minutes; and

d) a vent or check valve terminating said downstream flow control channel.

2. The microfluidic cartridge of claim 1, wherein said dual capillary stop comprises a first capillary stop positioned superiorly at the junction of said first intake channel with said staging union, said first capillary stop for controlledly gating entry of said first fluid into said reaction channel, and a second capillary stop positioned inferiorly at the junction of said second intake channel with said staging union, said second capillary stop for controlledly gating entry of said second fluid into said reaction channel, whereby wetting of said dual capillary stop initiates simultaneous parallel flow of said fluids as lamellae through said reaction channel, said first fluid on top of said second fluid, in response to said start signal.

3. The microfluidic cartridge of claim 1, wherein said start signal is generated by a pump configured for controlledly initiating fluid flow across said dual capillary stop, wherein said pump is fluidly connected to said downstream flow control channel, and initiates fluid flow by a suction stroke.

4. The microfluidic cartridge of claim 3, further comprising a throat segment fluidly interposed between said second end of said reaction channel and said runoff channel segment, wherein said throat segment is configured with a flow constrictor for resistively suppressing any pulsatile surge in said fluid flow through said reaction channel in response to said suction stroke.

5. The microfluidic cartridge of claim 1, wherein said reaction channel is configured with an upstream delay channel segment and a detection channel segment with detection window for detecting an agglutination reaction, and wherein said downstream flow control channel is configured with a volume selected to sustain laminar flow in said detection window at an apparent Reynolds Number between 0.1 and 10, as determined in said reaction channel for a duration of about thirty seconds to ten minutes, more preferably about thirty seconds to two minutes, said sustained laminar flow aiding detection.

6. The microfluidic cartridge of claim 1, wherein said body comprises a plurality of reaction channels with appended downstream flow control channels, operable in parallel, and terminating in a common vent, vented manifold, or check valve.

7. The microfluidic cartridge of claim 1, said first and second intake channels comprising first and second inlets, said first inlet for receiving said first fluid, said second inlet for receiving said second fluid.

8. The microfluidic cartridge of claim 1, wherein a dehydrated surfactant is deposited in said downstream flow control channel or in either or both said intake channels.

9. The microfluidic cartridge of claim 1, wherein one or more substrates of said body member are configured to modify the substrate surface energy by:

a) depositing a dehydrated surfactant in either or both said intake channels;

b) depositing a dehydrated surfactant in said downstream flow control channel;

c) passivating either or both said intake channels with a hydrophilic surface additive;

or, d) passivating said downstream flow control channel with a hydrophilic surface additive.

10. The microfluidic cartridge of claim 1, wherein said first fluid and said second fluid are selected from a biological sample and an immunological reagent, and said reaction channel with downstream flow control channel is configured for detecting an agglutination reaction between said biological sample and said immunological reagent.

11. The microfluidic cartridge of claim 10, wherein said immunological reagent is a dehydrated immunological reagent for rehydration at a point of use, wherein said dehydrated immunological reagent is deposited in an intake channel and wherein said dehydrated immunological reagent is optionally rehydrated with a hydrating reagent from a hydrating reagent reservoir or foil pouch formed within the body of the cartridge, said reservoir or foil pouch having a frangible seal and fluidic coupling to said intake channel.

12. The microfluidic cartridge of claim 1, further comprising a filter and plasma reservoir fluidly connected to one said intake channel, said filter for separating a plasma fraction from a whole blood sample.

13. A kit for performing a blood typing assay, which comprises said microfluidic cartridge of claim 10;

wherein said immunological reagent is selected from antibody, antigen, antibody to a blood group antigen, antigen to a blood group antibody, antibody to a protein, antibody to an antibody.

14. The kit of claim 13, wherein said microfluidic cartridge comprises a dehydrated immunological reagent and said kit comprises a rehydrating reagent solution.

15. The kit of claim 13, wherein said dehydrated immunological reagent is a dried matrix comprised of antigen- or antibody-coated latex particles.

16. The kit of claim 13, wherein said kit is configured for an blood typing assay selected from:

a) an agglutination assay for detecting a forward blood type;

b) an agglutination assay for detecting a reverse blood type;

or

c) an agglutination assay for detecting a positive direct or indirect Coombs test;

or

d) an agglutination assay for detecting a panel of blood types.

17. The kit of claim 13, wherein said kit is configured for testing a blood type selected from A, B, D, Lewis, MNS blood groups, P blood group, li blood group, Kell blood group, Kidd, Duffy blood group, Lutheran blood group, and Rhesus blood group in a whole blood sample or in packed red cells.

18. A kit for performing a crossmatch, which comprises a microfluidic cartridge of claim 1 configured for performing a crossmatch between a first blood sample from a prospective donor and a second sample from a transfusion recipient, wherein said first fluid comprises said blood sample, and said second fluid comprises said second blood sample, and said reaction channel is configured for detecting an agglutination reaction between a cell or cells of said first fluid and an antibody of said second fluid.

19. The kit of claim 18, wherein each said blood sample is selected from whole blood, washed red cells, packed red cells, platelets, serum, or plasma.

20. The kit of claim 19, wherein said plasma blood sample is formed at the point of use by filtration of whole blood through a filter into a plasma reservoir fluidly connected to said second intake channel, wherein said filter is formed in said cartridge body.

21. The kit of claim 18, further comprising a conglutinin reagent.

22. A kit for performing a Coombs test, which comprises said microfluidic cartridge of claim 1;

and further comprises an anti-human antibody reagent.

23. A method for performing an agglutination assay, which comprises:

a) introducing two flowing fluids into said reaction channel of a microfluidic cartridge of claim 1, wherein said cartridge is configured for forming a horizontally-stratified laminar fluid diffusion interface between said two flowing fluids; and

b) sustaining said horizontally-stratified laminar fluid diffusion interface in a laminar flow regime corresponding to a Reynolds Number of 0.1 to 10 for a duration of about thirty seconds to about ten minutes; and

c) detecting agglutination.

24. The microfluidic cartridge of claim 5, wherein the duration is about thirty seconds to two minutes.