Method for reducing the presence of amplification inhibitors in a reaction receptacle

• US 20060234263A1
• Filed: 03/10/2006
• Published: 10/19/2006
• Est. Priority Date: 03/10/2005
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Inventors: Light, James P. II
• Granted Patent: US 7,547,516 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• 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 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6851 Quantitative amplification
• CPC Class Codes: C12Q 2537/165 Mathematical modelling, e.g. logarithm, ratio
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 2021/6432 Quenching
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 2021/6441 with two or more labels
• CPC Class Codes: G01N 2035/00346 Heating or cooling arrangements
• CPC Class Codes: G01N 2035/00356 Holding samples at elevated temperature (incubation)
• CPC Class Codes: G01N 2035/00524 Mixing by agitating sample carrier
• CPC Class Codes: G01N 2035/0097 monitoring reactions as a function of time
• CPC Class Codes: G01N 2035/0441 for samples
• CPC Class Codes: G01N 2035/0443 for reagents
• CPC Class Codes: G01N 2035/0444 for cuvettes or reaction vessels
• CPC Class Codes: G01N 2035/0455 Coaxial carousels
• CPC Class Codes: G01N 2035/0491 Position sensing, encoding; closed-loop control
• CPC Class Codes: G01N 2035/103 using disposable tips
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6454 : using an integrated detector array
• CPC Class Codes: G01N 21/76 : Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 2201/0415 : Carrusel, sequential
• CPC Class Codes: G01N 35/00693 : Calibration
• CPC Class Codes: G01N 35/0092 : Scheduling
• CPC Class Codes: G01N 35/0098 : involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C
• CPC Class Codes: G01N 35/0099 : comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: G01N 35/025 : having a carousel or turntable for reaction cells or cuvettes
• CPC Class Codes: G01N 35/026 : having blocks or racks of reaction cells or cuvettes
• CPC Class Codes: G01N 35/028 : having reaction cells in the form of microtitration plates
• CPC Class Codes: G01N 35/04 : Details of the conveyor system G01N35/021 - G01N35/028 take precedence
• CPC Class Codes: G01N 35/1002 : Reagent dispensers
• CPC Class Codes: Y10S 435/808 : Optical sensing apparatus
• CPC Class Codes: Y10S 435/809 : Incubators or racks or holders for culture plates or containers
• CPC Class Codes: Y10T 436/113332 : with conveyance of sample along a test line in a container or rack
• CPC Class Codes: Y10T 436/25 : including sample preparation

Abstract

An automated analyzer for performing multiple diagnostic assays simultaneously includes multiple stations in which discrete aspects of the assay are performed on fluid samples contained in sample vessels. The analyzer includes stations for automatically preparing a sample, incubating the sample, preforming an analyte isolation procedure, ascertaining the presence of a target analyte, and analyzing the amount of a target analyte. An automated receptacle transporting system moves the sample vessels from one station to the next. A method for performing an automated diagnostic assay includes an automated process for isolating and amplifying a target analyte, and, in one embodiment, a method for real-time monitoring of the amplification process.

Claims

1. A method for reducing the presence of a nucleic acid amplification inhibitor in a reaction vessel, said method comprising:

(a) providing to a reaction vessel a surface treating agent and a nucleic acid containing material;

(b) agitating said reaction vessel to thereby mix the contents of said reaction vessel;

(c) exposing the contents of said reaction vessel to reagents and conditions sufficient to isolate a target nucleic acid, if present in said nucleic acid containing material, within said reaction vessel;

(d) during step (c), removing at least a portion of the contents of said reaction vessel;

(e) providing to said reaction vessel a wash solution;

(f) exposing the contents of said reaction vessel to reagents and conditions sufficient to isolate said target nucleic acid, if present in said nucleic acid containing material, within said reaction vessel;

(g) during step (f), removing at least a portion of the contents of said reaction vessel; and

(h) exposing the contents of said reaction vessel to reagents and conditions sufficient to amplify a target sequence present in said target nucleic acid,

wherein said surface treating agent is provided to said reaction vessel in an amount sufficient to reduce the presence of a nucleic acid amplification inhibitor during steps (d) and (g), and

wherein said surface treating agent is not an inhibitor of a nucleic acid amplification reaction.

2. The method of claim 1, wherein said surface treating agent is provided to said reaction vessel before said nucleic acid containing material is provided to said reaction vessel.

3. The method of claim 1, wherein said surface treating agent is provided to said reaction vessel after said nucleic acid containing material is provided to said reaction vessel.

4. The method of claim 1, wherein said surface treating agent prevents said inhibitor from adhering to an inner surface of said reaction vessel.

5. The method of claim 1, wherein said surface treating agent is not a detergent.

6. The method of claim 1, wherein said surface treating agent comprises silicone oil.

7. The method of claim 1 further comprising providing to said reaction vessel a solid support material for immobilizing said target nucleic acid.

8. The method of claim 7, wherein said solid support material comprises a magnetically responsive particle.

9. The method of claim 8, wherein the contents of said reaction vessel are exposed to a magnetic field during steps (c) and (f).

10. The method of claim 7, wherein said surface treating agent is provided to said reaction vessel before said solid support material is provided to said reaction vessel.

11. The method of claim 7, wherein said surface treating agent is provided to said reaction vessel after said solid support material is provided to said reaction vessel.

12. The method of claim 1, wherein said reaction vessel comprises a hydrophobic material.

13. The method of claim 12, wherein said reaction vessel comprises polypropylene.

14. The method of claim 1, wherein said inhibitor is provided by said nucleic acid containing material.

15. The method of claim 14, wherein said inhibitor is hemoglobin.

16. The method of claim 1, wherein said inhibitor is provided by said wash solution.

17. The method of claim 16, wherein said inhibitor is an anionic detergent.

18. The method of claim 1, wherein said reagents for amplifying said target sequence include a primer or promoter-primer, nucleoside triphosphates, and a nucleic acid polymerase capable of extending the 3′

end of said primer hybridized to said target nucleic acid.

19. A method for reducing the presence of a nucleic acid amplification inhibitor in a reaction vessel, said method comprising:

(a) removing at least a portion of a nucleic acid containing material from a reaction vessel;

(b) after step (a), providing to said reaction vessel a wash solution and a surface treating agent;

(c) agitating said reaction vessel to thereby mix the contents of said reaction vessel;

(d) exposing the contents of said reaction vessel to reagents and conditions sufficient to isolate a target nucleic acid, if present in said nucleic acid containing material, within said reaction vessel;

(e) during step (d), removing at least a portion of the contents of said reaction vessel; and

(f) exposing the contents of said reaction vessel to reagents and conditions sufficient to amplify a target sequence present in said target nucleic acid,

wherein said surface treating agent is provided to said reaction vessel in an amount sufficient to reduce the presence of a nucleic acid amplification inhibitor during step (e), and

wherein said surface treating agent is not an inhibitor of a nucleic acid amplification reaction.

20. The method of claim 19, wherein said wash solution and said surface treating agent are simultaneously provided to said reaction vessel.

21. The method of claim 19, wherein said wash solution and said surface treating agent are separately provided to said reaction vessel.

22. The method of claim 19, wherein said surface treating agent prevents said inhibitor from adhering to an inner surface of said reaction vessel.

23. The method of claim 19, wherein said surface treating agent is not a detergent.

24. The method of claim 19, wherein said surface treating agent comprises silicone oil.

25. The method of claim 19 further comprising providing to said reaction vessel a solid support material for immobilizing said target nucleic acid prior to step (a).

26. The method of claim 25, wherein said solid support material comprises a magnetically responsive particle.

27. The method of claim 26, wherein the contents of said reaction vessel are exposed to a magnetic field during step (d).

28. The method of claim 19, wherein said reaction vessel comprises a hydrophobic material.

29. The method of claim 28, wherein said reaction vessel comprises polypropylene.

30. The method of claim 19, wherein said inhibitor is provided by said nucleic acid containing material.

31. The method of claim 30, wherein said inhibitor is hemoglobin.

32. The method of claim 19, wherein said inhibitor is provided by said wash solution.

33. The method of claim 32, wherein said inhibitor is an anionic detergent.

34. The method of claim 19, wherein said reagents for amplifying said target sequence include a primer or promoter-primer, nucleoside triphosphates, and a nucleic acid polymerase capable of extending the 3′

end of said primer hybridized to said target nucleic acid.