Disposable test devices for performing nucleic acid amplification reactions

• US 6,410,275 B1
• Filed: 10/18/1999
• Issued: 06/25/2002
• 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: Cotter, Christopher, Graziano, Louis, Guy, Michel, Burg, Lawrence, Bishop, James Clement, Paris, Cécile, Wang, Thomas, Greer, James, Glassfold, Robert, Kluttz, Bryan, Tegeler, Garry, Morin, Michael, McKinley, Geoff, Colin, Bruno, Catanzariti, Luigi
• Primary Examiner(s): Redding, David A.
• Application Number:
  US09/420,139
• Time in Patent Office: 981 Days
• Field of Search: 435/91.1, 435/91.2, 435/91.5, 435/285.1, 435/287.2, 435/287.3, 435/287.6, 435/287.9, 435/288.2, 435/288.5, 435/286.1, 422/64, 422/101, 422/102, 935/78, 935/87, 935/88
• US Class Current: 435/91.1
• CPC Class Codes: B01L 2200/026 Fluid interfacing between devices or objects, e.g. connectors, inlet details
• 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 2400/049 vacuum
• 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: G01N 2035/00366 Several different temperatures used

Abstract

A unit dose test device for a nucleic acid amplification reaction has the form of a elongate disposable test strip. The test strip includes a dual-chamber reaction vessel pre-loaded with nucleic acid amplification reaction reagents, and a plurality of wells for processing a reaction occurring in the reaction vessel.

Claims

1. A test device for performing a nucleic acid amplification reaction, comprising:

a housing having a first end and a second end, said housing having a portion thereof defining a plurality of wells between said first and second ends;

said housing further comprising a dual chamber reaction vessel for a nucleic acid amplification reaction to be carried out therein, said dual chamber reaction vessel comprising a first chamber in which a first portion of a nucleic acid amplification reaction is performed, a second chamber in which a second portion of a nucleic acid amplification reaction is performed, an openable connecting conduit linking said first chamber to said second chamber and an externally actuated valve disposed in said openable connecting conduit, wherein an actuation of said valve opens said connecting conduit and allows fluid to flow through said connecting conduit from said first chamber to said second chamber.

2. The test device of claim 1, wherein said test device has a reagent inserted into said first chamber and a second reagent inserted into or placed in fluid communication with said second chamber, and wherein said test device further comprises a membrane affixed to said housing, said membrane applied to said housing to cover said first and second chambers and said wells after insertion of said first and second reagents into said test device.

3. The test device of claim 2, further comprising a cover member secured to said housing and overlying said membrane and said first and second chambers, wherein said cover member is moveable relative to said first chamber to thereby allow a user to access an opening in said membrane and introduce a sample into said first chamber.

4. The test device of claim 3, wherein said membrane comprises a first portion covering said first chamber and a second portion covering said second chamber and a third portion separating said first and second portions, wherein said cover member is moveable relative to said first chamber to allow a user to tear or perforate said third portion and thereby remove said first portion of said membrane from said test device.

5. The test device of claim 3, wherein said cover member further comprises:

a first portion, a hinge, and a second portion, said first portion covering said first chamber in a first position, said first portion moveable relative to said first chamber via said hinge to an elevated position enabling a user to access said first chamber, said second portion engaging said test device.

6. The test device of claim 5, wherein said first portion of said cover member further comprises an aperture in said first portion placed in registry with said first chamber when said first portion is in said first position, and wherein an absorbent mesh filter is placed within said aperture.

7. The test device of claim 1, wherein said test device further comprises at least one desiccant chamber for containing a desiccant, said desiccant chamber located on said test device in air or fluid communication with at least one of said first and second chambers, or both, of said dual chamber reaction vessel.

8. The test strip of claim 7, wherein said test device comprises two or more desiccant chambers for containing a desiccant, said two or more desiccant chambers located on said test device in air or fluid communication with at least one of said first and second chambers, or both, of said dual chamber reaction vessel.

9. The test device of claim 7, wherein said desiccant is in a pellet form.

10. The test device of claim 1, wherein a desiccant is loaded directly into at least one of said first and second chambers, or both, of said dual chamber reaction vessel.

11. The test device of claim 1, wherein said test device is made from a molded material and wherein a dessicant is molded into said test strip in the region of at least one of said first and second chambers.

12. The test device of claim 1, wherein said housing is in a form of an elongate strip with said wells spaced in an array along said strip, and wherein said second chamber is arranged in an aligned condition relation relative to said wells.

13. The test device of claim 1, wherein said test device further comprises an optical cuvette.

14. The test device of claim 1, wherein said dual chamber reaction vessel is integrally formed with said housing.

15. The test device of claim 1, wherein said housing comprises an aperture adjacent to said first end, and wherein said dual chamber reaction vessel comprises a separable piece inserted into said aperture.

16. The test device of claim 1, wherein said first and second chambers of said dual chamber reaction vessel further comprise a wall portion forming said first and second chambers having an exterior surface, and wherein said exterior surface of said wall portion is configured to come into touching contact with a heat sink having a predetermined form in a processing station for said test device.

34. The test device of claim 1, further comprising a cover member adapted for covering at least a portion of said housing, said cover member moveable relative to said housing so as to provide access to said dual chamber reaction vessel.

35. The test device of claim 34, wherein said cover member further comprises a button moveable relative to said housing having said cutting surface, said button operable to pierce a membrane applied to said housing in a manner to cover said dual chamber reaction vessel.

36. The test device of claim 34, wherein said cover member further comprises an aperture for receiving therein a porous filter, said cover mountable to said housing such that said porous filter is in registry with said first chamber of said dual chamber reaction vessel.

37. The test device of claim 34, wherein said cover member further comprises resilient snap means for releasably engaging said cover member to said housing.

17. A dual chamber disposable reaction vessel for a nucleic acid amplification reaction, comprising:

a first chamber for performing a first portion of a first nucleic acid amplification reaction,

a second chamber for performing a second portion of a nucleic acid amplification reaction,

a connecting conduit linking said first chamber to said second chamber,

a valve disposed in said connecting conduit controlling the flow of a fluid sample from said first chamber to said second chamber;

an enzyme pellet chamber disposed between said connecting conduit and said second chamber and adapted to contain an amplification enzyme pellet, such that said fluid sample passing from said first chamber to said second chamber passes through said enzyme pellet chamber; and

a membrane covering said first and second chambers.

18. The dual chamber disposable reaction vessel of claim 17, further comprising:

at least one desiccant chamber in communication with at least one of said first and second chambers, or both of said first and second chambers.

19. The dual chamber disposable reaction vessel of claim 17, wherein said dual chamber reaction vessel is contained in a disposable testing device comprising said dual chamber reaction vessel, at least one wash well, and a detection well.

20. The dual chamber reaction vessel of claim 17, wherein said dual chamber reaction vessel is contained in a disposable testing device comprising said dual chamber reaction vessel, and a decontamination well.

21. The dual chamber disposable reaction vessel of claim 19, wherein said testing device comprises an elongate strip.

22. The dual chamber disposable reaction vessel of claim 20, wherein said testing device comprises an elongate strip.

23. A method of constructing a unit dose nucleic acid amplification reaction testing apparatus, comprising the steps of:

forming a test device having (1) a dual chamber disposable reaction vessel comprising a first chamber, a second chamber, and a connecting conduit linking said first chamber with said second chamber, and (2) at least one intermediate processing well;

adding a first reagent to said first chamber;

adding a second reagent into said test device either in said second chamber or in an intermediate chamber in fluid communication with said second chamber; and

affixing a sealing membrane to said test device in a manner to cover said first and second chambers.

24. The method of claim 23, further comprising the step of affixing a cover member to said test device, said cover member moveable relative to first chamber to thereby allow access to said sealing film.

25. The method of claim 23, further comprising the step of affixing a base cap to said test device, said cap forming a channel between said first chamber and said connecting conduit.

26. The method of claim 23, further comprising the step of adding a desiccant into said test device in air communication with either said first reagent or said second reagent, or both said first and second reagent.

27. A method of performing a nucleic acid amplification reaction, comprising the steps of:

(1) providing a dual chamber reaction vessel ready for use in performing a nucleic acid amplification reaction, said vessel comprising a first reaction chamber, a second reaction chamber, a connecting conduit linking said first reaction chamber to said second reaction chamber and an externally actuated valve disposed in said connecting conduit, wherein an actuation of said valve opens said connecting conduit and allows fluid to flow through said connecting conduit from said first chamber to said second chamber;

comprising the steps of

placing a first reagent in fluid communication with said first reaction chamber and a second reagent in fluid communication with said second reaction chamber; and

sealing said first and second reaction chambers and said reagents from the environment; and

(2) conducting a nucleic acid amplification reaction in said vessel while maintaining a temperature differential between said first reaction chamber and the environment surrounding said second reagent, comprising the steps of;

introducing a sample into said first reaction chamber;

placing said first reaction chamber at a first temperature and conducting a first portion of said nucleic acid amplification reaction in said first reaction vessel at said first temperature;

actuating said valve and transferring a portion of the contents of said first reaction vessel to said second reaction chamber; and

placing said second reaction chamber at a second temperature different from said first temperature and conducting a second portion of said nucleic acid amplification reaction in said section reaction chamber at said second temperature.

28. The method of claim 27, wherein said dual chamber reaction vessel is embodied in a test strip having a plurality of wells arranged in array.

29. The method of claim 28, wherein said test strip has a major and minor axis having a predetermined length and width, respectively, said length and width selected such that said test strip is compatible with a detection instrument.

30. The method of claim 27, further comprising loading a desiccant into said dual chamber reaction vessel in air or fluid communication with at least one of said first and second reaction chambers, or both said first and second reaction chambers.

31. The method of claim 27, further comprising the step of affixing a cover to said dual chamber reaction vessel, wherein said cover is moveable relative to said first chamber to allow a user access to said first reaction chamber.

32. The method of claim 27, wherein said step of transferring comprises the step of drawing a vacuum on said dual chamber reaction vessel and drawing said at least a portion of the contents of said first chamber into said second chamber.

33. The method of claim 27, wherein said step of transferring comprises the step placing said dual chamber reaction vessel in a vacuum enclosure, applying vacuum to said vacuum enclosure, and then releasing said vacuum.