Nucleic acid amplification reaction station for disposable test devices

• US 6,429,007 B1
• Filed: 10/18/1999
• Issued: 08/06/2002
• Est. Priority Date: 05/02/1997
• Status: Expired due to Term

• 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, Gennari, Fabio, Kluttz, Bryan W., Dachaud, Jacques, McKinley, Geoff A., Guy, Michel, Jaravel, Cecile
• Primary Examiner(s): Redding, David A.
• Application Number:
  US09/420,140
• Time in Patent Office: 1,023 Days
• Field of Search: 435/286.5, 435/286.6, 435/287.2, 435/287.3, 435/287.6, 435/288.3, 435/288.4, 435/288.5, 422/63, 422/64, 422/68.1
• US Class Current: 435/286.5
• 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 station for conducting a nucleic acid amplification reaction in a disposable test device having a first reaction chamber containing a first nucleic acid amplification reagent and a second reaction chamber either containing or in fluid communication with a second nucleic acid amplification reagent, the station comprising:

a support structure receiving said test device;

at least one temperature control system for said test device, said temperature control system maintaining said first reaction chamber at a first temperature but simultaneously maintaining said second nucleic acid amplification reagent at a second temperature different from than said first temperature so as to preserve said second nucleic acid amplification reagent; and

an actuator operative on said test device to place said first and second reaction chambers in fluid communication with each other, said actuator operative on said test device after a reaction has occurred in said first reaction chamber at said first temperature;

whereby a second portion of said nucleic acid amplification reaction may occur in said second chamber with said second nucleic acid amplification reagent being preserved by maintenance of said second nucleic acid amplification reagent at said second temperature while said first chamber and said first nucleic acid amplification reagent are maintained at said first temperature.

2. The station of claim 1, further comprising a pneumatic system operative on said test device to draw a reaction solution from said first chamber to said second chamber after said actuator has operated on said device to place said first and second chambers in fluid communication with each other.

3. The station of claim 2, wherein said pneumatic system comprises a vacuum probe for said second chamber and a source of vacuum, wherein said source of vacuum draws said reaction solution from said first chamber into said second chamber.

4. The station of claim 2, wherein said pneumatic system comprises a vacuum enclosure and a source of vacuum,

said vacuum enclosure comprising a housing moveable with respect to said support structure and said test devices to thereby enclose said test devices and allow a vacuum to be drawn on said test devices,

said source of vacuum placed in communication with said vacuum enclosure and operative to withdraw air from said enclosure and admit air into said enclosure and thereby transfer said reaction solution from said first chamber to said second chamber.

5. The station of claim 1, wherein said test device comprises an elongate strip having (1) a reaction vessel comprising said first and second chambers and a connecting conduit linking said chambers, and (2) a plurality of wells.

6. The station of claim 1, wherein said support structure accommodates a plurality of said test devices and wherein said temperature control system and actuator are operative on each of said plurality of test devices, whereby said instrument is able to process said plurality of test devices substantially simultaneously.

7. The station of claim 1, wherein said station further comprises a mechanical agitation subsystem coupled to said support structure agitating said test devices after said test devices have been installed on said support structure.

8. The station of claim 1, wherein said station further comprises a front panel having a door providing access to said support structure, and wherein said support structure is coupled to a drive system for moving said support structure relative to said door between a retracted position and extended position, said extended position enabling a user to conveniently install said test devices on said support structure.

9. The station of claim 1, wherein said temperature control system comprises:

a first thermoelectric element installed in said instrument in proximity to a first portion of said support structure, wherein said first thermoelectric element maintains said first chamber at a first temperature, and

a second thermo-electric element installed in said instrument in proximity to a second portion of said support structure, wherein said second thermo-electric element maintains said second chamber at a second temperature.

10. The station of claim 9, further comprising first and second heat sinks in thermal contact with said first and second thermoelectric elements, respectively.

11. The station of claim 9, wherein said support structure comprises a pair of elongate groove features receiving a rim feature on said test device to thereby allow a technician to slide said test device relative to said support structure and thereby accurately positions said test device relative to said first and second thermoelectric elements.

31. The station of claim 1, wherein said test device has a form factor compatible with a separate, pre-existing analytic instrument for processing said test device after said nucleic acid amplification reaction has been performed.

12. An amplification station for conducting a nucleic acid amplification reaction in a plurality of disposable test devices, said test devices having a first reaction chamber containing a first nucleic acid amplification reagent and a second reaction chamber either containing or in fluid communication with a second nucleic acid amplification reagent, the amplification station comprising:

a support structure adapted to securely receiving a plurality of said test devices;

a temperature control system for said test devices, said temperature control system, for each of said test devices, maintaining said first reaction chamber at a first elevated temperature but simultaneously maintaining said second nucleic acid amplification reagent at a second temperature lower than said first temperature so as to preserve said second nucleic acid amplification reagent; and

an actuator assembly operative on each of said test devices to place said first and second reaction chambers in fluid communication with each other, said actuator assembly operative on said test devices after a reaction has occurred in said first reaction chamber at said first temperature;

whereby a second portion of said nucleic acid amplification reaction may occur in said second chamber with said second nucleic acid amplification reagent being preserved by maintenance of said second nucleic acid amplification reagent at said second temperature while said first chamber and said first nucleic acid amplification reagent are maintained at said first temperature.

13. The amplification station of claim 12, further comprising a pneumatic system operative on said test devices to draw a reaction solution from said first chamber to said second chamber after said actuator assembly has operated on said test devices to place said first and second chambers in fluid communication with each other.

14. The amplification station of claim 13, wherein said pneumatic system comprises a plurality of vacuum probes moveable into contact with said second chamber and a source of vacuum, wherein said source of vacuum draws said reaction solution from said first chamber into said second chamber when said plurality of vacuum probes has contacted said second chamber.

15. The amplification station of claim 13, wherein said pneumatic system comprises a vacuum enclosure and a source of vacuum,

said vacuum enclosure comprising a housing moveable with respect to said base, said housing covering said support structure and said test devices to thereby enclose said test devices and allow a vacuum to be drawn on said test devices,

said source of vacuum placed in communication with said vacuum enclosure and operative to withdraw air from said enclosure and admit air into said enclosure and thereby transfer said reaction solution from said first chamber to said second chamber.

16. The amplification station of claim 15, wherein said actuator assembly is fixedly mounted to said housing of said vacuum enclosure, and wherein said actuator assembly operates to open a connecting conduit linking said first reaction chamber to said second reaction chamber when said housing is lowered into contact with said base.

17. The amplification station of claim 16, wherein said actuator assembly comprises a plurality of forks, each of said forks operative on one of said test devices.

18. The amplification station of claim 12, wherein said test devices have the form of an elongate strip.

19. The amplification station of claim 12, further comprising a mechanical agitation system coupled to said support structure operative to agitate said plurality of test devices after said actuator assembly has opened said connecting conduit.

32. The amplification station of claim 12, wherein said test device has a form factor compatible with a separate, pre-existing analytic instrument for processing said test device after said nucleic acid amplification reaction has been performed.

20. 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 said test devices;

a temperature control system for said test devices, said temperature control system maintaining the temperature of said test devices according to a desired profiles(s) for said nucleic acid amplification reaction,

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

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

21. The amplification station of claim 20, wherein said pneumatic system comprises a plurality of vacuum probes moveable into contact with said second location of said test devices and a source of vacuum.

22. The amplification station of claim 21, wherein said pneumatic system comprises a vacuum enclosure and a source of vacuum,

said vacuum enclosure comprising a housing moveable with respect to said support structure, said housing covering said support structure and said test devices to thereby enclose said test devices and allow a vacuum to be drawn on said test devices,

said source of vacuum placed in communication with said vacuum enclosure and operative to withdraw air from said enclosure and admit air into said enclosure and thereby transfer said reaction solution from said first location to said second location.

23. The amplification station of claim 20, wherein said support structure is adapted to receive a plurality of said test devices and wherein said temperature control system and actuator assembly are operative on each of said plurality of test devices, whereby said instrument is able to process said plurality of test devices substantially simultaneously.

24. The amplification station of claim 23, wherein said amplification station further comprises a mechanical agitation subsystem coupled to said support structure agitating said test devices after said test devices have been installed on said support structure.

25. The amplification station of claim 20, wherein said instrument further comprises a front panel having a door providing access to said support structure, and wherein said support structure is coupled to a drive system for moving said support structure relative to said door between a retracted position and extended position, said extended position enabling a user to conveniently install said test devices on said support structure.

26. The amplification station of claim 20, wherein said temperature control system comprises:

a first thermo-electric element installed in said instrument in proximity to a first portion of said support structure, wherein said first thermo-electric element maintains said first chamber at a first temperature, and

a second thermoelectric element installed in said instrument in proximity to a second portion of said support structure, wherein said second thermoelectric element maintains said second chamber at a second temperature.

27. The amplification station of claim 26, further comprising first and second heat sinks in thermal contact with said first and second thermoelectric elements, respectively.

28. The amplification station of claim 20, wherein said test devices have the form of an elongate strip.

29. The amplification station of claim 28, wherein said elongate strip comprises a dual chamber reaction vessel having first and second reaction chambers linked together by a connecting conduit, and a plurality of wells arranged in a linear array.

30. The amplification station of claim 20, wherein said test device has a form factor compatible with a separate, pre-existing analytic instruent for processing said test device after said nucleic acid amplification reaction has been performed.

33. The amplification station of claim 20, wherein each of said test devices has a form factor compatible with a separate, pre-existing analytic instrument for processing said test device after said nucleic acid amplification reaction has been performed.

34. A station for conducting a nucleic acid amplification reaction in a disposable test device having a first reaction chamber containing a first nucleic acid amplification reagent and a second reaction chamber either containing or in fluid communication with a second nucleic acid amplification reagent, the station comprising:

a support structure receiving said test device;

a first temperature control system for said test device, said temperature control system maintaining said second reaction chamber at a predetermined temperature and preserving said second nucleic acid amplification reagent; and

an actuator operative on said test device to place said first and second reaction chambers in fluid communication with each other, said actuator operative on said test device after a reaction has occurred in said first reaction chamber.

35. The station of claim 34, further comprising a pneumatic system operative on said test device to draw a reaction solution from said first chamber to said second chamber after said actuator has operated on said device to place said first and second chambers in fluid communication with each other.

36. The station of claim 35, wherein said pneumatic system comprises a vacuum probe placed in contact with said second chamber and a source of vacuum, wherein said source of vacuum draws said reaction solution from said first chamber into said second chamber.

37. The station of claim 35, wherein said pneumatic system comprises a vacuum enclosure and a source of vacuum,

said vacuum enclosure comprising a housing moveable with respect to said support structure and said test devices to thereby enclose said test devices and allow a vacuum to be drawn on said test devices,

said source of vacuum placed in communication with said vacuum enclosure and operative to withdraw air from said enclosure and admit air into said enclosure and thereby transfer said reaction solution from said first chamber to said second chamber.

38. The station of claim 34, wherein said test device comprises an elongate strip having (1) a reaction vessel comprising said first and second chambers and a connecting conduit linking said chambers, and (2) a plurality of wells.

39. The station of claim 34, wherein said support structure accommodates a plurality of said test devices and wherein said temperature control system and actuator are operative on each of said plurality of test devices, whereby said instrument is able to process said plurality of test devices substantially simultaneously.

40. The station of claim 34, wherein said station further comprises a mechanical agitation subsystem coupled to said support structure agitating said test devices after said test devices have been installed on said support structure.

41. The station of claim 34, wherein said station further comprises a front panel having a door providing access to said support structure, and wherein said support structure is coupled to a drive system for moving said support structure relative to said door between a retracted position and extended position, said extended position enabling a user to conveniently install said test devices on said support structure.