Nucleic acid amplification reaction station for disposable test devices

• US 20110020921A1
• Filed: 09/24/2010
• Published: 01/27/2011
• Est. Priority Date: 05/02/1997
• Status: Active Grant

• Current Assignee: Biomerieux Incorporated (Compagnie Merieux Alliance SAS)
• Original Assignee: Biomerieux Incorporated (Compagnie Merieux Alliance SAS)
• Inventors: Colin, Bruno, Cotter, Christopher, Catanzariti, Luigi, Graziano, Louis, Paris, Cecile, Gennari, Fabio, Kluttz, Bryan W., Dachaud, Jacques, McKinley, Geoff A., Guy, Michel
• Granted Patent: US 9,562,260 B2
• Time in Patent Office: Days
• US Class Current: 435/287.2
• 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. An amplification station for conducting a plurality of nucleic acid amplification reactions in a plurality of unitary disposable test devices, the amplification station comprising:

a support structure adapted to securely receive a plurality of the test devices;

an actuator assembly operative on each of the test devices to open a fluid conduit in the test devices and thereby allow a reaction solution to flow from a first location in the test device to a second location in the test device, the first location having received a sample;

a pneumatic system operative on the test devices to draw a reaction solution from the first location to the second location after the actuator assembly has operated on the test devices to place the first and second portions in fluid communication with each other; and

a temperature control system for the test devices, the temperature control system maintaining the temperature of the second location in the test devices according to a desired profile or profiles for the nucleic acid amplification reaction.

2. The amplification station of claim 1, the test device further comprising a cover member.

3. The amplification station of claim 2, wherein the cover member is attached to the test device.

4. The amplification station of claim 1, wherein the test device further comprises a chamber in fluid communication with the second location, the chamber storing a unit dose of an enzymatic reagent.

5. The amplification station of claim 4, wherein the chamber is in fluid communication with the second location such that a reaction solution from the first location flows through the chamber en route to the second location.

6. The amplification station of claim 4, wherein the enzymatic reagent is in pellet form.

7. The amplification station of claim 1, wherein the test device further comprises a stored unit dose of primers.

8. The amplification station of claim 7, wherein the unit dose is in pellet form.

9. A disposable nucleic acid amplification test device comprising:

a body defining a first chamber for receiving a sample;

a conduit connecting the first chamber to a second chamber;

a nucleic acid amplification reaction enzyme in fluid communication with the second chamber;

a means disposed in the conduit for controlling the flow of the sample through the conduit; and

a device providing a barrier between the environment and the sample during the nucleic acid amplification of the sample.

10. The test device of claim 9, further comprising one or more chambers for wash operations on the sample.

11. The test device of claim 10, wherein the body is in the form of an elongate strip in which the first chamber, the second chamber and the one or more chambers for wash operations are in substantial alignment.

12. The test device of claim 9, further comprising a desiccant placed within the body in communication with the amplification enzyme.

13. The test device of claim 12, wherein the desiccant comprises one or more pellets of desiccant and a desiccant well for holding the one or more pellets.

14. The test device of claim 9, wherein the means for controlling the flow of the sample through the conduit into the second chamber comprises a valve, and wherein the valve is operated on to open the valve by an actuator external to the test device.

15. The test device of claim 9, further comprising reagents pre-loaded into the first chamber receiving the sample.

16. The test device of claim 15, further comprising a desiccant placed within the body in communication with the reagents.

17. The test device of claim 9, wherein the nucleic acid amplification reaction enzyme is placed within the second chamber.

18. The test device of claim 9, wherein the nucleic acid amplification reaction enzyme is placed within an intermediate chamber, the intermediate chamber connected to both the conduit and the second chamber.

19. The test device of claim 9, wherein the device providing the barrier is in the form of a sealing membrane.

20. The test device of claim 9, wherein the device providing the barrier is in the form of a cover member.