Automated nucleic acid assay system

• US 20030032172A1
• Filed: 07/02/2002
• Published: 02/13/2003
• Est. Priority Date: 07/06/2001
• Status: Abandoned Application

• 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: Colston, Billy W. Jr., Milanovich, Fred, Olson, Don, Belgrader, Phillip, Nasarabadi, Shanavaz L., Brown, Steve B., Marshall, Graham, Wolcott, Duane
• Application Number:
  US10/189,319
• Publication Number:
  US 20030032172A1
• Time in Patent Office: Days
• US Class Current: 435/287.2
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: G01N 1/28 Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Qmounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 ; laboratory gas handling apparatus B01L5/00

Abstract

A nucleic acid assay system includes a holding means that receives a sample and a reagent. A PCR reactor means amplifies the sample and produces an amplified sample. A detection means detects PCR amplicon. A transport means selectively transports the sample and the reagent relative to the holding means, the PCR reactor means, and the detection means. A control means is provided for selectively adding the reagent to the sample, mixing the sample and the reagent, performing PCR amplification, and detecting PCR amplicon. A decontamination means is provided for decontaminating the holding means, the PCR reactor means, and the detection means.

Claims

1. A nucleic acid assay system for analyzing a sample using a reagent, comprising:

holding means for receiving said sample and said reagent;

PCR reactor means for amplifying said sample;

detection means for detection of PCR amplicon;

transport means for selectively transporting said sample and said reagent to said holding means, said PCR reactor means, and said detection means, said transport means operatively connected to said holding means, said PCR reactor means, and said detection means;

control means for selectively adding said reagent to said sample, mixing said sample and said reagent, performing PCR amplification, and detecting PCR amplicon, said control means operatively connected to said holding means, said PCR reactor means, said detection means, and said transport means; and

means for decontaminating said holding means, said PCR reactor means, said detection means.

2. The nucleic acid assay system of claim 1, including conduits within said holding means, said PCR reactor means, said detection means, and said transport means;

and wherein said means for decontaminating said holding means, said PCR reactor means, said detection means includes means for decontaminating said conduits.

3. The nucleic acid assay system of claim 2, wherein said conduits include tubing.

4. The nucleic acid assay system of claim 2, wherein said conduits include microchannels.

5. The nucleic acid assay system of claim 2, wherein said conduits include passages within said PCR reactor means.

6. The nucleic acid assay system of claim 1, wherein said holding means mixes said sample with said reagent.

7. The nucleic acid assay system of claim 6, wherein said reagent is a PCR reagent.

8. The nucleic acid assay system of claim 6, wherein said PCR reagent includes primers.

9. The nucleic acid assay system of claim 6, wherein said PCR reagent includes oligos.

10. The nucleic acid assay system of claim 6, wherein said PCR reagent includes enzymes.

11. The nucleic acid assay system of claim 1, wherein said PCR reactor means cycles between a relatively high temperature and a relatively low temperature to produce PCR amplification.

12. The nucleic acid assay system of claim 1, wherein said PCR reactor means includes a section that can be held at a relatively high temperature and a section that can be held at a relatively low temperature and said PCR reactor means cycles said sample between said section that can be held at a relatively high temperature and said section that can be held at a relatively low temperature.

13. The nucleic acid assay system of claim 1, wherein said PCR reactor means includes an embedded thermocouple calibration conduit.

14. A nucleic acid assay method for analyzing a sample using a reagent, comprising the steps of:

providing a holding means for receiving said sample and said reagent;

providing a PCR reactor means for amplifying said sample;

providing a detection means for detection of PCR amplicon;

transporting said sample and said reagent to said holding means, said PCR reactor means, and said detection means;

said transport means operatively connected to said holding means, said PCR reactor means, and said detection means;

providing a decontamination means for decontaminating said holding means, said PCR reactor means, said detection means; and

providing a control means for selectively mixing said sample and said reagent, performing PCR amplification, detecting PCR amplicon, and decontaminating said holding means, said PCR reactor means, and said detection means;

said control means operatively connected to said holding means, said PCR reactor means, and said decontamination means.

15. The nucleic acid assay method of claim 14, including conduits within said holding means, said PCR reactor means, said detection means, and said transport means;

and wherein said decontamination means includes means for decontaminating said conduits.

16. The nucleic acid assay method of claim 15, wherein said conduits include tubing.

17. The nucleic acid assay method of claim 15, wherein said conduits include microchannels.

18. The nucleic acid assay method of claim 15, wherein said conduits include passages within said PCR reactor means.

19. The nucleic acid assay method of claim 14, wherein said sample and said reagent are mixed within said holding means.

20. The nucleic acid assay method of claim 19, wherein said reagent is a PCR reagent.

21. The nucleic acid assay method of claim 20, wherein said PCR reagent includes primers.

22. The nucleic acid assay method of claim 20, wherein said PCR reagent includes oligos.

23. The nucleic acid assay system of claim 20, wherein said PCR reagent includes enzymes.

24. The nucleic acid assay system of claim 14, including the step of cycling said sample between a relatively high temperature and a relatively low temperature to produce PCR amplification.

25. The nucleic acid assay system of claim 14, wherein said PCR reactor means includes a section that can be held at a relatively high temperature and a section that can be held at a relatively low temperature and said PCR reactor means cycles said sample between said section that can be held at a relatively high temperature and said section that can be held at a relatively low temperature.

26. A nucleic acid assay method for analyzing a sample, comprising the steps of:

utilizing a holding vessel for mixing said sample with a reagent;

utilizing a reactor for amplifying said sample and producing an amplified sample;

utilizing a detector for detecting PCR amplicon;

utilizing a fluidic system for selectively transporting said sample, said reagent, and said amplified sample relative to said holding means;

decontaminating and conditioning said nucleic acid assay system; and

utilizing a control for controlling the selectively adding of said reagent to said sample, mixing of said sample and said reagent, performing PCR amplification, detecting PCR amplicon, and decontaminating and conditioning said nucleic acid assay system.

27. The nucleic acid assay method of claim 26, wherein said reagent is a PCR reagent.

28. The nucleic acid assay method of claim 26, wherein said PCR reagent includes primers.

29. The nucleic acid assay method of claim 26, wherein said PCR reagent includes oligos.

30. The nucleic acid assay system of claim 26, wherein said PCR reagent includes enzymes.

31. The nucleic acid assay system of claim 26, including the step of cycling said sample between a relatively high temperature and a relatively low temperature to produce PCR amplification.