Nucleic acid amplification reaction station for disposable test devices

• US 7,807,449 B2
• Filed: 05/22/2006
• Issued: 10/05/2010
• Est. Priority Date: 05/02/1997
• Status: Expired due to Fees

• Current Assignee: Biomerieux Incorporated (Compagnie Merieux Alliance SAS)
• Original Assignee: Biomerieux Incorporated (Compagnie Merieux Alliance SAS)
• Inventors: Paris, Cecile, Catazariti, Luigi, Dachaud, Jacque, Colin, Bruno, Cotter, Christopher, Graziano, Louis, Gennari, Fabio, Kluttz, Bryan W., McKinley, Geoff A., Guy, Michel
• Primary Examiner(s): BOWERS, NATHAN ANDREW
• Application Number:
  US11/438,938
• Publication Number:
  US 20060263871A1
• Time in Patent Office: 1,597 Days
• Field of Search: 435/286.5, 435/286.6, 435/287.2, 435/287.3, 435/288.3, 435/288.4, 435/288.5, 435/305.2, 435/288.7, 422/102, 422/103
• US Class Current: 435/287.3
• CPC Class Codes: B01L 2200/026 Fluid interfacing between devices or objects, e.g. connectors, inlet details
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0809 rectangular shaped
• CPC Class Codes: B01L 2300/0825 Test strips
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 2400/0633 with moving parts
• CPC Class Codes: B01L 2400/0644 rotary valves
• CPC Class Codes: B01L 2400/0655 pinch valves
• 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 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: C12Q 1/6848 characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
• CPC Class Codes: G01N 2035/0436 with pre-packaged reagents, i.e. test-packs
• CPC Class Codes: G01N 35/026 having blocks or racks of reaction cells or cuvettes

Abstract

An instrument for conducting nucleic acid amplification reactions in a disposable test device. The test device includes a first reaction chamber containing a first nucleic acid amplification reagent (e.g., primers and nucleotides) and a second reaction chamber either containing, or in fluid communication, with a second nucleic acid amplification reagent (e.g., an amplification enzyme such as RT). The instrument includes a support structure receiving the test device. A temperature control system maintains the first reaction chamber at a first elevated temperature but simultaneously maintains the second nucleic acid amplification reagent at a second temperature lower than the first temperature so as to preserve the second nucleic acid amplification reagent. An actuator operates on a fluid conduit in the test device to place the first and second reaction chambers in fluid communication with each other after a reaction has occurred in the first reaction chamber at the first temperature. A pneumatic system is also provided that assists in fluid transfer of a reaction solution from the first chamber to the second chamber.

Claims

1. A nucleic acid reaction processing station for processing nucleic acid reactions occurring in a plurality of disposable test devices having a nucleic acid sample contained therein and comprising a first reaction chamber and a second reaction chamber and a valve connecting the first reaction chamber to the second reaction chamber, said station comprising

a) a plurality of individual disposable test device receiving locations, each of said locations adapted for receiving an individual one of said disposable test devices;

b) a plurality of actuators, one actuator per disposable test device, operative on the valve of said disposable test devices to open fluid communication between said first reaction chamber and said second reaction chamber;

c) a pneumatic system including a source of vacuum connected to a vacuum enclosure moveable into a position to cover the disposable test devices and form a vacuum chamber for said disposable test devices, wherein the plurality of actuators are positioned within the vacuum enclosure, the pneumatic system operative to transfer a reaction solution contained in the first reaction chamber to the second reaction chamber after opening of the fluid communication between the first reaction chamber and the second reaction chamber;

d) at least one thermal module comprising

1) a thermo-electric heat source,

2) a first thermal transfer structure positioned adjacent to at least one of said plurality of individual disposable test device receiving locations so as to be in direct thermal contact with said first reaction chamber of a disposable test device received in said disposable test device receiving location, and

3) a second thermal transfer structure positioned adjacent to at least one of said plurality of individual disposable test device receiving locations so as to be in direct thermal contact with said second reaction chamber of a disposable test device received in said disposable test device receiving location,

e) a temperature control system operative of said thermo-electric heat source so as to produce a desired temperature profile to said first thermal transfer structure and thereby a first desired temperature profile to said first reaction chamber and a second desired temperature profile to said second thermal transfer structure and thereby a second desired temperature profile to said second reaction chamber, in accordance with a nucleic acid reaction to be performed on a sample contained in the disposable test device; and

f) an optical reader assembly comprising an optical sensor positioned to detect a disposable test device received within an individual disposable test device receiving location.

2. The amplification station of claim 1, wherein said disposable test device further comprises an enzyme pellet well in fluid communication with said second reaction chamber.

3. The station of claim 2, wherein the pneumatic system is operative to draw a reaction solution from said first reaction chamber through said enzyme pellet well to said second reaction chamber after said actuator has operated on the valve of said disposable test device to thereby deliver an enzyme contained in the enzyme pellet well into said second reaction chamber.

4. The station of claim 1, wherein said pneumatic system further comprises a drive mechanism for reciprocating said vacuum enclosure between upper and lower positions relative to said plurality of individual disposable test device receiving locations.

5. The station of claim 1, wherein said station comprises at least four separate individual disposable test device receiving locations, each of which is in direct thermal contact with said first and second thermal transfer structures.

6. The station of claim 5, wherein said station comprises at least four separate optical sensors positioned to detect a disposable test device received within each individual disposable test device receiving location.

7. A nucleic acid reaction processing station for processing nucleic acid reactions occurring in a plurality of disposable test devices having a nucleic acid sample contained therein and comprising a first reaction chamber, a second reaction chamber, chamber containing an enzyme, and a valve connecting the first reaction chamber to the chamber containing the enzyme and the second reaction chamber, said station comprising

a) a plurality of individual disposable test device receiving locations, each of said locations adapted for receiving an individual one of said disposable devices;

b) a plurality of actuators, one per test device, operative on the valve of said disposable test devices to open fluid communication between said first reaction chamber, said second reaction chamber and said chamber containing an enzyme;

c) a pneumatic system including a source of vacuum connected to a vacuum enclosure moveable into a position to cover the disposable test devices and form a vacuum chamber for said disposable test devices, wherein the plurality of actuators are positioned within the vacuum enclosure, the pneumatic system operative to transfer a reaction solution contained in the first reaction chamber to the chamber containing the enzyme and the second reaction chamber after actuation of the valve;

d) at least one thermal module comprising

1) a thermo-electric heat source, and

2) at least one thermal transfer structure positioned adjacent to at least one of said plurality of individual disposable test device receiving locations so as to be in direct thermal contact with said second reaction chamber of a disposable test device received in said disposable test device receiving location,

e) a temperature control system operative of said thermo-electric heat source so as to produce a desired temperature profile to said at least one thermal transfer structure and thereby a desired temperature profile to said second reaction chamber of said disposable test device in accordance with a nucleic acid reaction to be performed on a sample contained in the disposable test device; and

f) an optical reader assembly comprising an optical sensor positioned to detect a disposable test device received within an individual disposable test device receiving location.

8. The station of claim 7, wherein the pneumatic system is operative to draw a reaction solution from said chamber containing an enzyme to said second reaction chamber after said actuator has operated on said valve to place said first and second reaction chambers and said chamber containing an enzyme in fluid communication with each other to thereby deliver an enzyme contained in the enzyme chamber into said second reaction chamber.

9. The station of claim 7, wherein said pneumatic system further comprises a drive mechanism for reciprocating said vacuum enclosure between upper and lower positions relative to said plurality of individual disposable test device receiving locations.

10. The station of claim 7, wherein said station comprises at least four separate individual disposable test device receiving locations, each of which. is in direct thermal contact with said at least one thermal transfer structure.

11. The station of claim 10, wherein said station comprises at least four separate optical sensors positioned to detect a disposable test device received within each individual disposable test device receiving location.