Continous flow thermal device

• US 20050282206A1
• Filed: 06/10/2005
• Published: 12/22/2005
• Est. Priority Date: 08/16/2001
• Status: Active Grant

• Current Assignee: Qiagen GmbH (Qiagen NV)
• Original Assignee: Corbett Research Pty Limited (Qiagen NV)
• Inventors: Michael Corbett, John, Corbett, John Michael Jr.
• Granted Patent: US 7,709,250 B2
• Time in Patent Office: Days
• US Class Current: 435/6.12
• CPC Class Codes: B01L 2200/026 Fluid interfacing between devices or objects, e.g. connectors, inlet details
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2300/1838 using fluid heat transfer medium
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: G01N 2021/7786 Fluorescence
• CPC Class Codes: G01N 21/78 producing a change of colour
• CPC Class Codes: G01N 35/08 using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis

Abstract

The present invention provides a thermal cycling device including: a heat exchanger body (3) having a longitudinal axis and longitudinally divided to provide at least two segments which are able to be heated to different temperatures so that said body has peripheral surfaces of different temperatures; a conduit (2) extending about said body so as to be in thermal contact with said peripheral surfaces; a first delivery device to deliver a first fluid to said conduit to cause said fluid to pass therealong and therefore change in temperature as the fluid passes said segments; and a second delivery device to deliver a second fluid to said conduit so as to flow with said first liquid and therefore also change in temperature. The fluid then leaves the device by conduit (4). The invention also is directed to an injector means for use in the thermal cycling device of this invention, comprising: a septum (26); a needle (20) which is able to pierce the septum; a reservoir (27) in fluid contact with septum (26) having an inlet (28) and an outlet (30); wherein when in use, the needle is urged through the fluid in said reservoir and then through the septum while continually purging the reservoir with said purge fluid. The present invention also includes a scanning detector (5) fitted to the outside of the heat exchange body (3) which monitors the course of reaction inside the conduit (2).

Claims

1. A thermal cycling device including:

a heat exchanger body having a longitudinal axis and longitudinally divided to provide at least two segments which are able to be heated to different temperatures so that said body has peripheral surfaces of different temperatures;

a conduit extending about said body so as to be in thermal contact with said peripheral surfaces;

a first delivery device to deliver a first fluid to said conduit to cause said fluid to pass therealong and therefore change in temperature as the fluid passes said segments; and

a second delivery device to deliver a second fluid to said conduit so as to flow with said first liquid and therefore also change in temperature.

2. The device according to claim 1 wherein the body is divided into 3 segments able to be heated to different temperatures.

3. The device according to claim 2 wherein the circumferential proportion of the segments is 1:

2;

1.

4. The device according to claim 3 wherein the temperatures in each segment are:

(a) about 94°

C., (b) about 55°

C., and (c) 72°

C. respectively.

5. The device according to claim 4 wherein the first fluid comprises a sample to be analysed and reagents used in the analysis and the second fluid comprises a transport fluid.

6. The device according to claim 5 wherein a sample to be analysed is separated from a following sample to be analysed by a carrier fluid to substantially prevent contamination between said samples.

7. The device according to claim 6 wherein the carrier fluid is silicon oil, PCR grade water or any synthetic oil which is devoid of RNA or DNA.

8. The device according to claim 7 wherein the sample to be analysed comprises at least one nucleic acid.

9. The device according to claim 8 wherein the analysis is the polymerase chain reaction or the ligase chain reaction.

19. The device according to any one of claims 1 to 18 wherein the body is a regular, three dimensional shaped body which can accommodate conduit to effectively transfer heat to a fluid in the conduit.

20. The device according to claim 19 wherein the body is cylindrically shaped.

21. The device according to any one of claims 1 to 20 wherein the body has a groove in the surface to accommodate the conduit in thermal contact with said surface.

22. The device according to claims 21 wherein said conduit extends angularly about said body relative to said axis.

23. The device according to claim 22, where said conduit extends about said body in a series of consecutive windings.

24. The device according to claim 23 wherein the number of windings is between about 35 and about 60.

25. The device according to claim 24 wherein the number of windings is about 60.

26. The device according to any one of claims 1 to 25 wherein the conduit is tubing which is able to conduct heat from the heat exchanger body to fluid in the conduit so that a selectable change in temperature in a segment of the heat exchanger body accurately causes a selectable change in temperature in a particular portion of the conduit adjacent that portion of the heat exchanger body.

27. The device according to claim 26 wherein the interior surface of the conduit is hydrophobic.

28. The device according to claim 27 wherein the conduit is PTFE tubing or a similar tubing.

31. The device according to any one of claims 1 to 30 wherein a back pressure is applied at the end of the fluid stream.

32. The device according to claim 31 wherein the back pressure is between about 300 and about 700 kPa.

33. The device according to any one of claims 1 to 32 wherein the body is hollow.

34. The device according to any one of claims 1 to 32 wherein the body is solid.

35. The device according to any one of claims 1 to 34 comprising heating and/or cooling means to achieve a selectable temperature.

36. The device according to claim 35 wherein the heating device is a Peltier device.

41. The device according to any one of claims 1 to 36 including the injector means according to any one of claims 37 to 40.

42. The device according to claim 41 wherein the first fluid is delivered by the injector means.

43. The device according to claim 42 wherein a marker reagent for monitoring a chemical reaction in said conduit is added to at least one of said first and second fluids.

44. The device according to claim 43 wherein said marker device is a fluorescent dye, a coupling reagent where a change in colour indicates progress of a reaction, or a chromogenic substrate.

45. The device according to any one of claims 1 to 36 and 41 to 44 further comprising a scanning detector, wherein said scanning detector is able to monitor the course of a reaction occurring in said conduit.

46. The device according to claim 45, wherein said scanning detector is able to move past successive windings of said conduit, thereby to detect the course of a reaction in the conduit by scanning an individual winding of said conduit and analysing.

47. The device according to claim 45 wherein the heat exchanger body is able to move relative to the scanning detector, thereby to detect the course of a reaction in the conduit by scanning an individual winding of said conduit and analysing.

48. A method of performing a polymerase chain reaction or ligase chain reaction, according to any one of claims 1 to 9, 19 to 36, and 41 to 47.

51. A nucleic acid prepared by using the device according to any one of claims 1 to 9, 19 to 36 and 41 to 36;

or the method according to claim 48.

10. A thermal cycling device including:

a heat exchanger body having a longitudinal axis and transversely divided to provide at least two segments which are able to be heated to different temperatures so that said body has peripheral surfaces of different temperatures;

a conduit extending about said body so as to be in thermal contact with said peripheral surfaces;

a first delivery device to deliver a first fluid to said conduit to cause said fluid to pass therealong and therefore change in temperature as the fluid passes said segments; and

a second delivery device to deliver a second fluid to said conduit so as to flow with said first liquid and therefore also change in temperature.

11. The device according to claim 10 wherein a temperature gradient is applied across the at least two segments.

12. The device according to claim 11 wherein the temperature gradient is from about 50°

C. to about 950 C.

13. The device according to claim 12 wherein the first fluid comprises a sample to be analysed and the second fluid comprises a transport fluid.

14. The device according to claim 13 wherein the sample comprises a nucleic acid.

15. The device according to claim 14 wherein the analysis is a nucleic acid melt detection assay.

29. The device according to any one of claims 5 to 9, 13 to 15 and 17 to 28 wherein the transport fluid is delivered under pressure.

30. The device according to claim 29 wherein the pressure is from about 700 to about 7,000 kPa.

49. A method of performing a nucleic acid melt detection assay comprising using the device according to any one of claims 10 to 15, 19 to 36, and 41 to 47.

16. An isothermal device including:

a heat exchanger body having a longitudinal axis and a peripheral surface and means for maintaining the temperature of said peripheral surface at a constant selectable temperature;

a conduit extending about said body so as to be in thermal contact with said peripheral surface;

a first delivery device to deliver a first fluid to said conduit to cause said fluid to pass therealong and therefore change in temperature as the fluid passes said peripheral surface; and

a second delivery device to deliver a second fluid to said conduit so as to flow with said first liquid and therefore also change in temperature.

17. The device according to claim 16 wherein the first fluid comprises an antibody and an antigen to said antibody and the second fluid comprises a transport fluid.

18. The device according to claim 16 or claim 17 used to analyse an antibody-antigen binding reaction.

50. A method of analysing an antibody-antigen binding reaction comprising using the device according to any one of claims 16 to 36, and 41 to 47.

37. An injector means for use in a thermal cycling device comprising:

a septum;

a needle which is able to pierce the septum;

a reservoir in fluid contact with the septum, having an inlet and an outlet, and able to accommodate a purge fluid which is able to enter the reservoir via the inlet and leave the reservoir via the outlet;

wherein when in use, the needle passes through fluid in said reservoir and then through the septum.

38. The injector means according to claim 37 wherein the purge fluid is silicon oil, PCR grade water or any synthetic oil which is devoid of RNA or DNA.

39. The injector means according to claim 37 or claim 38 wherein the septum is made of material which closes about the hole made by a previous penetration.

40. The injector means according to any one of claims 37 to 39 wherein the septum is made of rubber.