Polymerase chain reaction system

• US 20020191826A1
• Filed: 12/29/2000
• Published: 12/19/2002
• Est. Priority Date: 01/04/2000
• Status: Active Grant

• Current Assignee: Lawrence Livermore National Security LLC (Government of the United States of America)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Belgrader, Phil, Meyer, Peter L., Benett, William J., Milanovich, Fred P., Hadley, Dean R., Stratton, Paul L., Richards, James B.
• Granted Patent: US 6,699,713 B2
• Time in Patent Office: Days
• US Class Current: 382/129
• 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: C12M 1/34 Measuring or testing with condition measuring or sensing means, e.g. colony counters

Abstract

A portable polymerase chain reaction DNA amplification and detection system includes one or more chamber modules. Each module supports a duplex assay of a biological sample. Each module has two parallel interrogation ports with a linear optical system. The system is capable of being handheld.

Claims

1. A polymerase chain reaction DNA amplification and detection system, comprising:

one or more sample chamber modules adapted to contain biological sample volumes,

each of said sample chamber modules having the ability to support a duplex assay, and

parallel interrogation ports operatively connected to said sample chamber modules.

2. the polymerase chain reaction DNA amplification and detection system of claim 1, including a linear detection system operatively connected to each of said interrogation ports.

3. the polymerase chain reaction DNA amplification and detection system of claim 1, including a linear detection systems operatively connected to said interrogation ports, wherein said linear detection systems are positioned substantially parallel to each other.

4. The polymerase chain reaction DNA amplification and detection system of claim 1, including a main body with said one or more sample chamber modules removeably connected to said main body.

5. The polymerase chain reaction DNA amplification and detection system of claim 4, wherein said main body and said one or more sample chamber modules are capable of being handheld.

6. The polymerase chain reaction DNA amplification and detection system of claim 4, wherein said main body is less than about 12 inches long.

7. The polymerase chain reaction DNA amplification and detection system of claim 4, wherein said main body is about 10 inches or less long.

8. The polymerase chain reaction DNA amplification and detection system of claim 4, wherein said main body is less than about 6 inches wide.

9. The polymerase chain reaction DNA amplification and detection system of claim 4, wherein said main body is about 4 inches or less wide.

10. The polymerase chain reaction DNA amplification and detection system of claim 4, wherein said main body is less than about 4 inches deep.

11. The polymerase chain reaction DNA amplification and detection system of claim 4, wherein said main body is about 2 inches or less deep.

12. The polymerase chain reaction DNA amplification and detection system of claim 4, wherein said main body and said one or more sample chamber modules weigh less than about 5 pounds.

13. The polymerase chain reaction DNA amplification and detection system of claim 4, wherein said main body and said one or more sample chamber modules weigh a bout 2 pounds or less.

14. The polymerase chain reaction DNA amplification and detection system of claim 4, wherein said main body and said one or more sample chamber modules are battery operated.

15. The polymerase chain reaction DNA amplification and detection system of claim 1, wherein each of said interrogation ports has a unique optical fluorescence detection system

16. The polymerase chain reaction DNA amplification and detection system of claim 1, wherein each of said parallel interrogation ports has its own optical fluorescence detection system.

17. The polymerase chain reaction DNA amplification and detection system of claim 1, wherein each of said sample chamber modules has multiple ports which interrogate distinct regions within said sample volume using its own optical fluorescence detection system.

18. The polymerase chain reaction DNA amplification and detection system of claim 1, including a sleeve reaction chamber having a plurality of grooves in the sleeve for reducing thermal mass and increasing surface area of the sleeve.

19. The polymerase chain reaction DNA amplification and detection system of claim 18, wherein said plurality of grooves includes at least one longitudinally extending groove, and at least one radially extending groove, and at least one radially extending groove connected to said longitudinally extending groove to form a passageway there-through.

20. The polymerase chain reaction DNA amplification and detection system of claim 19, wherein said plurality of grooves includes at least one longitudinally extending groove on opposite sides of said sleeve, and at least one radially extending groove connected to each of said longitudinally extending grooves to form passageways there-through.

21. The polymerase chain reaction DNA amplification and detection system of claim 20, wherein two longitudinally extending grooves are located on opposite sides of said sleeve, and wherein said plurality of connecting radially extending grooves are located along opposite end sections of said sleeve.

22. The polymerase chain reaction DNA amplification and detection system of claim 21, wherein each of said grooves is of a V-shaped configuration.

23. The polymerase chain reaction DNA amplification and detection system of claim 22, wherein said sleeve is constructed of silicon.

24. The polymerase chain reaction DNA amplification and detection system of claim 23, wherein said sleeve reaction chamber is composed of two sections, said sections being bonded or clamped together.

25. The polymerase chain reaction DNA amplification and detection system of claim 24, wherein said two sections are configured to define a chamber to enclose an insert therebetween such that said insert is in direct contract with at least a portion of walls of said two sections defining the chamber.

26. The polymerase chain reaction DNA amplification and detection system of claim 1, wherein an LED directs light energy into each of said parallel interrogation ports.

27. The polymerase chain reaction DNA amplification and detection system of claim 26, including bandpass filters in each of said parallel interrogation ports.

28. The polymerase chain reaction DNA amplification and detection system of claim 1, including a non-orthogonal fluorescence, highly compact optical detection system.

29. The polymerase chain reaction DNA amplification and detection system of claim 1, including an assay status user interface.

30. The polymerase chain reaction DNA amplification and detection system of claim 1, including one or more master microprocessors and one or more slave microcontrollers.

31. The polymerase chain reaction DNA amplification and detection system of claim 1, including a positive calling algorithm for determining when a biological agent was present in said sample volume.

32. The polymerase chain reaction DNA amplification and detection system of claim 1, including a status display that indicates the viability of a particular assay.

33. The polymerase chain reaction DNA amplification and detection system of claim 25, including electronics located in said main body and forced cooling ducts for efficiently coupling external air through said sleeve reaction chamber then out of said main body, thereby cooling said sample and said electronics simultaneously.

35. The method of polymerase chain reaction DNA amplification and detection of claim 34, wherein said light energy is directed into said biological sample through a linear detection system.

36. The method of polymerase chain reaction DNA amplification and detection of claim 34, including the steps of:

heating said sample using a heating unit in a silicon base, and cooling said sample.

37. The method of polymerase chain reaction DNA amplification and detection of claim 36, including forcing cooling air through a sleeve reaction chamber to cooling said biological sample.

38. The method of polymerase chain reaction DNA amplification and detection of claim 34, wherein said sample chamber module includes an individual sample chamber and including the step of temporarily overheating said sample chamber for the purpose of having said biological sample achieve and hold a desired temperature as rapidly as possible.

39. The method of polymerase chain reaction DNA amplification and detection of claim 34, including the step of:

determining that an assay of said sample is positive based on the relationship between a multiplicity of signal increases and a per-cycle threshold value for said increases.

40. The method of polymerase chain reaction DNA amplification and detection of claim 39, including the step of:

displaying in real-time the degree of certainty or of reliability for each said assay.

34. A method of polymerase chain reaction DNA amplification and detection, comprising the steps of:

placing a biological sample in a sample chamber module that has the ability to support a duplex assay, and

directing light energy into said biological sample through parallel interrogation ports operatively connected to said sample chamber module.