Device for treating nucleic acids from a sample

• US 5,746,978 A
• Filed: 07/17/1997
• Issued: 05/05/1998
• Est. Priority Date: 06/15/1994
• Status: Expired due to Term

• Current Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Original Assignee: Boehringer Mannheim GmbH (Roche Holding AG)
• Inventors: Bienhaus, Gerhard, Lange, Hans
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): REES, DIANE
• Application Number:
  US08/895,920
• Time in Patent Office: 292 Days
• Field of Search: 435/6, 435/5, 435/91.2, 435/814, 435/815, 422/68.1, 536/25.3, 417/243, 209/11, 210/222, 210/500.1, 210/143, 210/175, 210/294, 423/210, 55/220
• US Class Current: 422/68.1
• 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: C12Q 1/6848 characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
• CPC Class Codes: Y10S 435/814 Enzyme separation or purification
• CPC Class Codes: Y10S 435/815 by sorption

Abstract

Device for the treatment of nucleic acids from the sample, comprising a first reaction chamber for separating the nucleic acids from other sample components, and a second reaction chamber for the amplification of the nucleic acids, connected to said first reaction chamber via a controllable transport path.

Claims

1. A device for treating nucleic acids in a sample solution, said device comprising:

a first reaction chamber for separating nucleic acids from other sample components in the sample solution;

a second reaction chamber for the amplification of purified nucleic acids which have been conveyed to the second reaction chamber by means of a transport path.

a transport path connecting said first reaction chamber to the second reaction chamber;

a controlling means connected to the transport path for controlling liquid flow through the transport path into the second reaction chamber wherein said controlling means allows nucleic acids separated from other sample components in the sample solution to be transported into the second reaction chamber and prevents liquid containing said other sample components from being transported into the second reaction chamber thereby conveying purified nucleic acids to said second reaction chamber.

2. A device according to claim 1, wherein said first reaction chamber, said second reaction chamber, and said transport path are configured to be a closed system during a treatment of nucleic acids therein, said closed system including at least one selectively openable opening in one of said first reaction chamber nd said second reaction chamber, for adding reagents thereto.

3. A device according to claim 1, wherein said second reaction chamber includes an opening for removing a reaction mixture therefrom.

4. A device as recited in claim 1, wherein said first reaction chamber includes immobilization means for immobilizing nucleic acids therein.

5. A device according to claim 1, wherein said transport path comprises three-way distribution means for distributing substances along three paths therein, the first path leading from said three-way distribution means to the first reaction chamber, the second path leading from the three-way distribution means to the second reaction chamber, and the third path leading from the three-way distribution means to a waste chamber.

6. A device as recited in claim 1, further comprising heating means for heating at least one of the first and second reaction chambers.

7. A device as recited in claim 1, wherein said second reaction chamber comprises an opening therein for the addition of reagents, said reagents being used for an amplification reaction, said second reaction chamber also having closing means for selectively closing said opening.

8. A device as recited in claim 2, wherein at least said first reaction chamber and said second reaction chamber comprise removable and replaceable elements.

9. A device according to claim 1, wherein said first reaction chamber and said second reaction chamber are of a modular design and can be assembled and disassembled, such that a plurality of first reaction chambers and second reaction chambers can be modularly configured to provide an apparatus which can be used as a multi-step reaction apparatus.

10. A device according to claim 1, wherein at least one of said first reaction chamber and said second reaction chamber can be emptied by the application of a vacuum thereto.

11. A device as recited in claim 1, wherein atleast one of said first reaction chamber and said second reaction chamber includes magnetic separation means for magnetically separating magnetic phases from non-magnetic phases of said sample.

12. A device as recited in claim 4, wherein said apparatus includes a plurality of first reaction chambers and second reaction chambers, for a multiplicity of applications.

13. A system for the treatment of nucleic acids obtained from a sample solution said system comprising:

pipetting means for pipetting liquids therethrough;

receiving means coupled to said pipetting means for receiving a plurality of the devices of claim 1.

14. A system as recited in claim 13, further comprising heating means for heating at least one of said plurality of devices.

15. A system as recited in claim 13, further comprising at least one magnet positioned adjacent to at least one of said plurality of devices.

18. A method for treating nucleic acids in a sample, said method comprising:

providing a device according to claim 1 and placing the sample solution into the first reaction chamber of said device wherein nucleic acids in said sample solution are concentrated and separated from other sample components in the sample solution thereby concentrating and purifying said nucleic acids;

transporting said concentrated and purified nucleic acids to the second reaction chamber through the transport path by operating said controlling means; and

amplifying said purified nucleic acids which have been conveyed to the second reaction chamber by means of the transport path.

21. A device according to claim 1, wherein said controlling means comprises a valve.

22. A device according to claim 1, wherein said controlling means also controls an enrichment of nucleic acids by controlling an amount of nucleic acid containing solution flowing into the second reaction chamber.

23. A device according to claim 1, wherein said controlling means also controls the amplification of the nucleic acids by controlling an amount of nucleic acid containing solution flowing into the second reaction chamber.

24. A device according to claim 21, wherein said valve opens when a vacuum pressure is applied to the first reaction chamber.

25. A system according to claim 13, further comprising a controlling means coupled to said transport means for said controlled transport of fluid.

26. A device according to claim 23, wherein said controlling means controls an amplification of nucleic acids in at least one reaction chamber of the plurality of reaction chambers.

27. A device according to claim 26, wherein said controlling means also controls an enrichment of the sample being transported through the transport means by controlling an amount of nucleic acid containing solution flowing into the second reaction chamber.

28. A system according to claim 25, wherein said control means comprises a valve.

31. A method according to claim 18, wherein said providing step includes a substep of providing a controlling means for controlling a concentration of nucleic acids in the sample.

33. A method for treating nucleic acids in a sample, said method comprising:

providing a device according to claim 1;

placing a sample containing nucleic acids into the first reaction chamber of said device wherein said sample is liquefied in the first reaction chamber;

transporting said liquefied sample to the second reaction chamber of the device through the transport path by operating the controlling means; and

combining said nucleic acids of said liquefied sample with nucleic acid amplifying reagents which have been placed in the second reaction chamber; and

amplifying said nucleic acids.

34. A method as recited in claim 33, wherein said separation step comprises retaining said nucleic acids from said sample in a filer material.

35. A method as recited in claim 33, wherein said separation step comprises non-specific immobilization of said nucleic acids to an adsorbent contained in said first reaction chamber.

36. A method as recited in claim 33, wherein said separation step comprises biospecific immobilization of said nucleic acids to biospecific adsorbents.

37. A method as recited in claim 33, wherein said step of liquefying said sample includes a step of separating nucleic acids from substance interfering with a subsequent samplification of said nucleic acids, thereby producing the prepared sample.

38. A method for treating nucleic acids in a sample, said method comprising:

providing a device according to claim 1;

placing a sample containing nucleic acids into the first reaction chamber of said device wherein nucleic acids are separated from substances in the solution that inhibit nucleic acid amplification;

transporting said separated nucleic acids in the sample solution to the second reaction chamber of the device through the transport path by operating the controlling means; and

combining said separated nucleic acids with nucleic acid amplifying reagents which have been placed in the second reaction chamber; and

amplifying said purified nucleic acids.

39. A method as recited in claim 38, wherein said step of separating nucleic acids includes liquefying the sample, thereby producing the prepared sample.

40. A method as recited in claim 38, wherein said separation step comprises retaining said nucleic acids from said sample in a filter material.

41. A method as recited in claim 39, wherein said separation step comprises nonspecific immobilization of said nucleic acids to an adsorbent contained in said first reaction chamber.

42. A method as recited in claim 38, wherein said separation step comprises biospecific immobilization of said nucleic acids to biospecific adsorbents.

16. A module for treatment of nucleic acid containing fluids, said module comprising:

A treatment chamber for separating nucleic acids from other sample components in a sample;

a closure coupled to said treatment chamber which is configured to be open and closed from the outside of said chamber by applying a vacuum to said closure, said closure enclosing and sealing said treatment chamber when closed.

17. A module as recited in claim 16, wherein said closure comprises a valve, and said valve is mounted to a bottom of said treatment chamber.

32. A module as recited in claim 16, wherein said closure controls a treatment of the sample in the treatment chamber.

19. A device for treating nucleic acids in a sample solution, said device comprising:

a first reaction means for separating nucleic acids from other sample components in the sample solution;

a second reaction means for the amplification of purified nucleic acids which have been conveyed to the second reaction chamber by means of a transport path;

a transport path connecting said first reaction means to the second reaction means;

a controlling means connected to the transport path for controlling liquid flow through the transport path into the second reaction means wherein said controlling means allows nucleic acids separated from other sample components in the sample solution to be transported into the second reaction means and prevents liquid containing said other sample components from being transported into the second reaction means thereby conveying purified nucleic acids to said second reaction means.

20. A device according to claim 19, further comprising at least one selective sealing means for selectively sealing a system formed by the first reaction means, the second reaction means, and the transport means.

29. A device according to claim 19, further comprising controlling means coupled to said transport means, for controlling the sample being transported through the transport means.

30. A device according to claim 29, wherein said control means comprises a valve.