Assay cartridges and methods of using the same

Patent metadata

Field	Value
Publication number	US-9731297-B2
Application number	US-201213343834-A
Country	US
Kind code	B2
Filing date	Jan 5, 2012
Priority date	Jan 6, 2011
Publication date	Aug 15, 2017
Grant date	Aug 15, 2017

Abstract

Assay cartridges are described that have purification, reaction, and detection zones and other fluidic components which can include sample chambers, waste chambers, conduits, vents, reagent chambers, reconstitution chambers and the like. The assay cartridges are used to conduct multiplexed nucleic acid measurements. Also described are kits including such cartridges, methods of using the same, and a reader configured to analyze an assay conducted using an assay cartridge.

First claim

What is claimed is:

1. An assay cartridge for conducting a PCR analysis of a sample, said cartridge comprising a fluidic network including
(a) a plurality of air vent ports arranged as one or two rows of apertures on the cartridge,
(b) a primary flow path comprising, from a proximal to a distal end, an inlet, a purification zone, a PCR reaction zone, and a detection zone, wherein said primary flow path further comprises one or more of said air vent ports,
(c) one or more chambers connected to an additional one of said air vent port, and
(d) one or more fluidic conduits each intersecting said primary flow path and fluidically connected to said chamber, wherein a fluidic conduit of said one or more fluidic conduits comprises a multi-conduit fluidic junction including (i) a first conduit connecting said primary flow path and said chamber, and (ii) a second conduit connecting said chamber to said additional air vent port, wherein said fluidic conduit is configured to be in communication with an optical fluid sensor at a position distal from said fluidic junction and a metered volume of fluid is defined by the distance between said fluidic junction and said distal position,
wherein said PCR reaction zone comprises a first temperature controlled zone and a second temperature controlled zone, and a restriction zone positioned between said first and second temperature controlled zones, and said fluidic network is configured to shuttle a metered volume of fluid between said first and second temperature controlled zones during a PCR reaction conducted in said PCR reaction zone.

2. The assay cartridge of claim 1 wherein said assay cartridge further comprises a plurality of chambers selected from a sample chamber, a reagent chamber, a PCR reagent chamber, a reconstitution chamber, a waste chamber, and a mixing chamber.

3. The assay cartridge of claim 2 wherein said assay cartridge further comprises a sample chamber, a plurality of reagent chambers, a plurality of PCR reagent chambers, a plurality of reconstitution chambers, a waste chamber, and a mixing chamber.

4. The assay cartridge of claim 3 wherein said primary flow path is intersected by a sample chamber multi-conduit fluidic junction including (i) a first sample chamber conduit connecting said primary flow path and said sample chamber; and (ii) a second sample chamber conduit connecting said sample chamber to a sample chamber air vent port, wherein said sample chamber fluidic junction is in communication with a sample chamber optical fluid sensor at a position distal from said sample chamber fluidic junction and said metered volume of fluid is defined by the distance between said sample chamber fluidic junction and said distal position.

5. The assay cartridge of claim 4 wherein said primary flow path is intersected by a mixing chamber multi-conduit fluidic junction including (i) a first mixing chamber conduit connecting said primary flow path and said mixing chamber; and (ii) a second mixing chamber conduit connecting said mixing chamber to a mixing chamber air vent port, wherein said mixing chamber fluidic junction is in communication with a mixing chamber optical fluid sensor at a position distal from said mixing chamber fluidic junction and said metered volume of fluid is defined by the distance between said mixing chamber fluidic junction and said distal position.

6. The assay cartridge of claim 5 wherein said primary flow path further comprises a series of Z-transitions at a position distal from said mixing chamber fluidic junction.

7. The assay cartridge of claim 1 wherein said purification zone is fluidically connected to a first waste chamber and a purification reagent chamber.

8. The assay cartridge of claim 7 wherein said purification zone comprises, from a proximal to a distal end,
(i) a purification multi-conduit fluidic junction including
(a) a first purification reagent chamber conduit connecting said primary flow path and said purification reagent chamber; and
(b) a second purification reagent chamber conduit connecting said purification reagent chamber and a purification reagent chamber air vent port;

(ii) an integrated purification membrane positioned in said primary flow path of said purification zone; and
(iii) a waste multi-conduit fluidic junction including
(a) a first waste chamber conduit connecting said primary flow path and said waste chamber; and
(b) a second waste chamber conduit connecting said waste chamber and a waste chamber air vent port.

9. The assay cartridge of claim 8 wherein said purification zone comprises an integrated purification membrane positioned on a support frit.

10. The assay cartridge of claim 8 wherein said integrated purification membrane is compressed prior to cartridge assembly.

11. The assay cartridge of claim 8 wherein said integrated purification membrane comprises a glass fiber membrane.

12. The assay cartridge of claim 8 wherein said membrane has a retention volume of less than 10 uL.

13. The assay cartridge of claim 8 wherein said membrane has a retention volume of less than 5 uL.

14. The assay cartridge of claim 8 wherein said membrane has a retention volume of less than 2 uL.

15. The assay cartridge of claim 8 wherein said purification zone further comprises a pre-heating zone preceding said integrated purification membrane.

16. The assay cartridge of claim 8 wherein said purification zone comprises an additional temperature controlled zone.

17. The assay cartridge of claim 1 wherein said primary flow path is intersected at said reaction zone by one or more reagent multi-conduit junctions connecting (i) said primary flow path and one or more reaction reagent chambers; and (ii) said one or more reagent chambers and one or more reagent chamber air vent ports.

18. The assay cartridge of claim 17 wherein said assay cartridge further comprises one or more reconstitution chambers and said one or more reagent multi-conduit junctions further connect (iii) said primary flow path and said one or more reconstitution chambers at said reaction zone.

19. The assay cartridge of claim 17 wherein said reaction zone comprises a reaction zone inlet and a reaction zone outlet, and said one or more multi-conduit junctions intersect said reaction zone at said reaction zone inlet and/or outlet.

20. The assay cartridge of claim 1 wherein said primary flow path is intersected at said detection zone by a detection reagent multi-conduit junction connecting (i) said primary flow path and one or more detection reagent chambers; and (ii) said one or more detection reagent chambers and one or more detection reagent air vent ports.

21. The assay cartridge of claim 20 wherein said detection zone comprises a set of immobilized reagents distributed in said detection zone along said primary flow path.

22. The assay cartridge of claim 21 wherein said detection zone further comprises an exposed working electrode on which said reagents are immobilized.

23. The assay cartridge of claim 20 wherein said detection zone is configured to maintain uniform fluid flow throughout said detection zone.

24. The assay cartridge of claim 20 wherein said detection zone comprises a square geometry.

25. The assay cartridge of claim 20 wherein said primary flow path is U-shaped within said detection zone.

26. The assay cartridge of claim 1 wherein said detection zone further comprises a detection temperature controlled z

Assignees

• Glezer Eli N
• Higgins Stephen
• Kovacs Sandor
• Kumar Sudeep
• Page Kenneth
• Roth Kristian
• Sigal George
• Meso Scale Technologies Llc

Inventors

• Glezer Eli N
• Higgins Stephen
• Kovacs Sandor
• Kumar Sudeep
• Page Kenneth
• Roth Kristian
• Sigal George

Classifications

• B01L2200/0605

  Metering of fluids · CPC title

• B01L3/5027

  by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip · CPC title

• B01L7/525 Primary

  with physical movement of samples between temperature zones · CPC title

• B01L2200/16

  Reagents, handling or storing thereof · CPC title

• B01L2200/0621

  Control of the sequence of chambers filled or emptied · CPC title