Reagent cartridge and methods for detection of cells

• US 9,540,675 B2
• Filed: 02/09/2015
• Issued: 01/10/2017
• Est. Priority Date: 10/29/2013
• Status: Active Grant

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: GeneWEAVE BioSciences, Inc. (Roche Holding AG)
• Inventors: De Forest, Nikol, Frei, Werner, Rey, Diego, Roy, Shaunak, Shukla, Soni, Griswold, Ryan C., Olson, Kenneth G., Richardson, Bruce J., Yee, Victor H.
• Primary Examiner(s): EPPS -SMITH, JANET L
• Application Number:
  US14/617,631
• Publication Number:
  US 20150218613A1
• Time in Patent Office: 701 Days
• Field of Search: 435/287.1, 435/5
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/042 Caps; Plugs
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/047 Additional chamber, reservoir
• CPC Class Codes: B01L 2300/049 Valves integrated in closure
• CPC Class Codes: B01L 2300/0672 Integrated piercing tool
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0478 pistons
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0683 mechanically breaking a wall or membrane within a channel or chamber
• CPC Class Codes: B01L 3/502 with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• CPC Class Codes: B01L 3/50825 Closing or opening means, corks, bungs closures for containers B65D; means for removing stoppers B67B7/02
• CPC Class Codes: B01L 3/523 with means for closing or opening
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: C12Q 1/18 Testing for antimicrobial activity of a material
• CPC Class Codes: C12Q 1/6897 involving reporter genes operably linked to promoters
• CPC Class Codes: G01N 2021/0325 Cells for testing reactions, e.g. containing reagents
• CPC Class Codes: G01N 2021/0328 Arrangement of two or more cells having different functions for the measurement of reactions
• CPC Class Codes: G01N 2021/6443 Fluorimetric titration
• CPC Class Codes: G01N 2021/7786 Fluorescence
• CPC Class Codes: G01N 21/6486 : Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 21/76 : Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 21/77 : by observing the effect on a chemical indicator

Abstract

An apparatus includes a housing and an actuator. The housing, which defines a reagent volume that can receive a reagent container, can be removably coupled to a reaction chamber. The housing includes a puncturer that defines a transfer pathway in fluid communication with the reagent volume. A delivery portion of the housing defines a delivery pathway between the transfer pathway and the reaction chamber when the housing is coupled to the reaction chamber. The actuator has a plunger portion disposed within the reagent volume. An engagement portion of the actuator can be manipulated to move the plunger portion within the reagent volume to deform the reagent container. The puncturer can pierce a frangible portion of the reagent container to convey a reagent from the reagent container into the reaction chamber via the transfer pathway and/or the delivery pathway.

Claims

1. An apparatus, comprising:

a housing configured to be removably coupled to a reaction chamber, the housing defining a reagent volume configured to receive a reagent container, the housing including a puncturer defining a transfer pathway in fluid communication with the reagent volume, the puncturer and the housing being monolithically constructed, the housing including a delivery portion defining a delivery pathway between the transfer pathway and the reaction chamber when the housing is coupled to the reaction chamber; and

an actuator having a plunger portion disposed within the reagent volume, an engagement portion of the actuator configured to be manipulated to move the plunger portion within the reagent volume to deform the reagent container, the puncturer configured to pierce a frangible portion of the reagent container to convey a reagent from the reagent container into the reaction chamber via the transfer pathway and the delivery pathway.

2. The apparatus of claim 1, wherein the transfer pathway is perpendicular to the frangible portion of the reagent container.

3. The apparatus of claim 1, wherein the puncturer defines a plurality of transfer pathways in fluid communication with the reagent volume and the delivery pathway, the transfer pathway being one of the plurality of transfer pathways.

4. The apparatus of claim 1, wherein a portion of the delivery pathway partially surrounds the puncturer.

5. The apparatus of claim 1, wherein an exit portion of the delivery pathway defines an exit axis that intersects a sidewall of the reaction chamber.

6. The apparatus of claim 1, further comprising:

the reagent container disposed within the reagent volume, the reagent container including a skirt surrounding the frangible portion, the skirt and a surface of the delivery portion forming a substantially fluid-tight seal; and

a lock member disposed within the reagent volume, the lock member configured to limit movement of the skirt relative to the housing when the reagent container is deformed to maintain the substantially fluid-tight seal between the skirt and the surface.

7. The apparatus of claim 1, further comprising:

the reagent container disposed within the reagent volume, the reagent container including a skirt surrounding the frangible portion, a first portion of the skirt and a first surface of the delivery portion forming a substantially fluid-tight seal, a second portion of the skirt configured to engage a retention portion of the housing to limit movement of the skirt relative to the housing when the reagent container is deformed, the retention portion within the reagent volume.

8. The apparatus of claim 1, wherein the plunger portion of the actuator and a portion of the housing collectively define a seal to fluidically isolate the reagent volume from a volume outside of the housing.

9. The apparatus of claim 1, further comprising:

the reagent container disposed within the reagent volume, the reagent container containing a plurality of transduction particles.

10. The apparatus of claim 9, wherein the plurality of transduction particles is engineered to be incapable of lytic replication.

11. The apparatus of claim 1, further comprising:

the reagent container disposed within the reagent volume, the reagent container containing the reagent formulated to react with a plurality of reporter molecules in a sample to enhance production of a signal.

12. The apparatus of claim 1, further comprising:

the reagent container disposed within the reagent volume, the reagent container containing at least one of a nutrient, an antibiotic, a lysis reagent or a sterilizing reagent.

13. The apparatus of claim 1, further comprising:

the reagent container disposed within the reagent volume, the reagent container containing tridecanal.

14. The apparatus of claim 1, wherein the reagent is a first reagent, the apparatus further comprising:

the reaction chamber containing a second reagent formulated to react with a sample within the reaction chamber.

15. An apparatus, comprising:

a housing configured to be coupled to a reaction chamber, the housing defining a reagent volume configured to receive a reagent container, the housing including a puncturer having a sharp point configured to pierce a frangible portion of the reagent container, the puncturer defining a plurality of transfer pathways spaced circumferentially about the sharp point, each transfer pathway from the plurality of transfer pathways in fluid communication with an inner volume of the reaction chamber when the housing is coupled to the reaction chamber; and

an actuator having a plunger portion disposed within the reagent volume, an engagement portion of the actuator configured to be manipulated to move the plunger portion within the reagent volume such that the sharp point pierces the frangible portion of the reagent container to convey a reagent from the reagent container into the reaction chamber via the plurality of transfer pathways.

16. The apparatus of claim 15, wherein the puncturer and the housing are monolithically constructed.

17. The apparatus of claim 15, wherein the plurality of transfer pathways produces a discontinuous cross-sectional shape within the puncturer at a cross-sectional position below the sharp point.

18. The apparatus of claim 15, wherein each transfer pathway from the plurality of transfer pathways is separate from the other transfer pathways from the plurality of transfer pathways.

19. The apparatus of claim 15, wherein each transfer pathway from the plurality of transfer pathways is spaced an equidistance from the sharp point.

20. The apparatus of claim 19, wherein the plurality of transfer pathways includes at least four separate transfer pathways.

21. The apparatus of claim 15, wherein the housing includes a delivery portion defining a delivery pathway that places the plurality of transfer pathways in fluid communication with the inner volume of the reaction chamber when the housing is coupled to the reaction chamber.

22. The apparatus of claim 21, wherein a portion of the delivery pathway partially surrounds the sharp point of the puncturer.

23. The apparatus of claim 21, wherein the reagent is a first reagent, the apparatus further comprising:

the reagent container disposed within the reagent volume, the reagent container containing the first reagent formulated to react with a plurality of reporter molecules in a sample to enhance production of a signal; and

the reaction chamber containing a second reagent formulated to react with the sample to limit production of the signal.

24. The apparatus of claim 23, wherein the second reagent contains an antibiotic in a solid form, the antibiotic formulated to limit production of the plurality of reporter molecules.

25. The apparatus of claim 21, further comprising:

the reagent container disposed within the reagent volume, the reagent container including a skirt surrounding the frangible portion, the skirt and a surface of the delivery portion forming a substantially fluid-tight seal; and

a lock member configured to limit movement of the skirt relative to the housing when the sharp point pierces the frangible portion of the reagent container to maintain the substantially fluid-tight seal between the skirt and the surface.

26. An apparatus, comprising:

a housing configured to be coupled to a reaction chamber, the housing defining a reagent volume configured to receive a reagent container, the housing including a puncturer configured to pierce a frangible portion of the reagent container, the puncturer defining a plurality of transfer pathways spaced apart along an outer surface of the puncturer, each transfer pathway from the plurality of transfer pathways in fluid communication with an inner volume of the reaction chamber when the housing is coupled to the reaction chamber; and

an actuator having a plunger portion disposed within the reagent volume, an engagement portion of the actuator configured to be manipulated to move the plunger portion within the reagent volume such that the puncturer pierces the frangible portion of the reagent container to convey a reagent from the reagent container into the reaction chamber via the plurality of transfer pathways.

27. The apparatus of claim 26, wherein each of the plurality of transfer pathways is perpendicular to the frangible portion of the reagent container.

28. The apparatus of claim 26, wherein the plurality of transfer pathways produces a discontinuous cross-sectional shape within the outer surface of the puncturer at a cross-sectional position below a sharp point of the puncturer.

29. The apparatus of claim 26, wherein each transfer pathway from the plurality of transfer pathways is separate from the other transfer pathways from the plurality of transfer pathways.

30. The apparatus of claim 26, wherein the plurality of transfer pathways includes at least four separate transfer pathways.

31. The apparatus of claim 26, wherein the housing includes a delivery portion defining a delivery pathway that places the plurality of transfer pathways in fluid communication with the inner volume of the reaction chamber when the housing is coupled to the reaction chamber.

32. The apparatus of claim 31, wherein a portion of the delivery pathway partially surrounds a sharp point of the puncturer.

33. The apparatus of claim 26, wherein the reagent is a first reagent, the apparatus further comprising:

the reagent container disposed within the reagent volume, the reagent container containing the first reagent formulated to react with a plurality of reporter molecules in a sample to enhance production of a signal; and

the reaction chamber containing a second reagent formulated to react with the sample to limit production of the signal.

34. The apparatus of claim 33, wherein the second reagent contains an antibiotic in a solid form, the antibiotic formulated to limit production of the plurality of reporter molecules.

35. The apparatus of claim 26, wherein the housing includes a threaded portion configured to be matingly coupled to a threaded portion of the reaction chamber.

36. The apparatus of claim 26, wherein the puncturer and the housing are monolithically constructed.