Automated process for detecting the presence of a target nucleic acid in a sample

• US 7,482,143 B2
• Filed: 06/29/2005
• Issued: 01/27/2009
• Est. Priority Date: 05/01/1998
• Status: Expired due to Fees

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Inventors: Burns, Ralph E., Nieglos, Donald J., Kling, John E., Jakub, Cheryl A., Horner, Glenn A., Smith, Robert J., Schneider, Robert E., Ammann, Kelly G., Hansberry, Ernest V.
• Primary Examiner(s): LU, FRANK WEI MIN
• Application Number:
  US11/169,112
• Publication Number:
  US 20050239127A1
• Time in Patent Office: 1,308 Days
• Field of Search: 435/6, 435/91.1, 435/91.2, 435/183, 435/284.4, 435/287.1, 435/287.2, 436/94, 436/501, 536/23.1, 536/24.3, 536/24.33, 536/25.3, 536/25.32
• US Class Current: 435/91.2
• CPC Class Codes: B01F 29/10 with receptacles rotated about two different axes, e.g. receptacles having planetary motion
• CPC Class Codes: B01F 29/30 Mixing the contents of individual packages or containers, e.g. by rotating tins or bottles
• CPC Class Codes: B01F 29/322 of two or more containers supported for simultaneous mixing, e.g. for bottles in crates
• CPC Class Codes: B01F 29/40354 arranged for planetary motion
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• 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: B01L 7/5255 by moving sample containers
• CPC Class Codes: B01L 9/06 Test-tube stands; Test-tube holders
• CPC Class Codes: B03C 1/282 with associated accumulation indicator, e.g. Hall sensor
• CPC Class Codes: B03C 1/288 disposed at the outer circumference of a recipient
• CPC Class Codes: B03C 1/30 Combinations with other devices, not otherwise provided for
• CPC Class Codes: B03C 2201/26 for use in medical applications
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6832 Enhancement of hybridisation reaction
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 2537/10 the purpose or use of
• 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 2035/0437 : Cleaning cuvettes or reaction vessels
• CPC Class Codes: G01N 2035/0455 : Coaxial carousels
• CPC Class Codes: G01N 2035/103 : using disposable tips
• 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 35/1065 : Multiple transfer devices
• CPC Class Codes: Y10T 156/1105 : Delaminating process responsive to feed or shape at delamination
• CPC Class Codes: Y10T 156/1906 : Delaminating means responsive to feed or shape at delamination
• CPC Class Codes: Y10T 29/53657 : to apply or remove a resilient article [e.g., tube, sleeve, etc.]
• CPC Class Codes: Y10T 29/53683 : Spreading parts apart or separating them from face to face engagement
• CPC Class Codes: Y10T 29/53843 : Tube, sleeve, or ferrule inserting or removing
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/111666 : Utilizing a centrifuge or compartmented rotor
• CPC Class Codes: Y10T 436/113332 : with conveyance of sample along a test line in a container or rack
• CPC Class Codes: Y10T 436/114998 : with treatment or replacement of aspirator element [e.g., cleaning, etc.]
• CPC Class Codes: Y10T 436/119163 : with aspirator of claimed structure
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

An automated process for detecting the presence of a target nucleic acid in a sample. The process is performed in a stand-alone unit and includes separating the target nucleic acid from other material present in the sample, engaging a robotic pipettor to combine the separated target nucleic acid and reagents for performing an amplification reaction, and performing an amplification reaction. A product of the amplification reaction is detected as an indication of the presence of the target nucleic acid in the sample.

Claims

1. A process for detecting the presence of a target nucleic acid in a sample containing the target nucleic acid, the process comprising the automated steps of:

a) immobilizing the target nucleic acid on a magnetically responsive particle;

b) subjecting the magnetically responsive particle to a magnetic field and aspirating non-immobilized components of the sample;

c) combining the target nucleic acid immobilized in step a) and reagents for performing an amplification reaction using a robotic pipettor;

d) performing an amplification reaction which produces an amplification product indicative of the presence of the target nucleic acid in the sample; and

e) detecting the amplification product of step d) as an indication of the presence of the target nucleic acid in the sample,

wherein the steps of the process are performed in a self-contained analyzer.

2. The process of claim 1, wherein the non-immobilized components of the sample include non-target nucleic acid.

3. The process of claim 1, wherein the target nucleic acid is hybridized to a capture probe prior to immobilizing the target nucleic acid on the magnetically responsive particle.

4. The process of claim 1 further comprising the step of forming a complex comprising the amplification product and a probe, wherein the amplification product is detected in step e) by detecting the formation of the complex.

5. The process of claim 4, wherein the probe includes a detectable label.

6. The process of claim 4, wherein the amplification product is a nucleic acid sequence that is the same as or complementary to a sequence of the target nucleic acid, and wherein the probe is hybridized to the amplification product.

7. The process of claim 1, wherein steps a)-e) are performed at a plurality of stations contained within a housing of the analyzer.

8. The process of claim 7, wherein the stations are located on a processing deck.

9. The process of claim 7, wherein the analyzer is movable.

10. The process of claim 1, wherein the amplification reaction is selected from the group consisting of a polymerase chain reaction, a strand displacement amplification, and a transcription-associated amplification.

11. A process for detecting the presence of a target nucleic acid in a sample containing the target nucleic acid, the process comprising the automated steps of:

a) immobilizing the target nucleic acid on a magnetically responsive particle;

b) subjecting the magnetically responsive particle to a magnetic field and aspirating non-immobilized components of the sample;

c) after step b), washing the magnetically responsive particle one or more times with a wash solution;

d) combining the target nucleic acid immobilized in step a) and reagents for performing an amplification reaction using a robotic pipettor;

e) performing an amplification reaction which produces an amplification product indicative of the presence of the target nucleic acid in the sample; and

f) detecting the amplification product of step e) as an indication of the presence of the target nucleic acid in the sample,

wherein the steps of the process are performed in a self-contained analyzer.

12. The process of claim 11 further comprising the step of forming a complex comprising the amplification product and a probe, wherein the amplification product is detected in step f) by detecting the formation of the complex.

13. The process of claim 12, wherein the probe includes a detectable label.

14. The process of claim 11, wherein steps a)-f) are performed at a plurality of stations contained within a housing of the analyzer.

15. The process of claim 14, wherein the stations are located on a processing deck.

16. The process of claim 14, wherein the analyzer is movable.

17. The process of claim 11, wherein the amplification reaction is selected from the group consisting of a polymerase chain reaction, a strand displacement amplification, and a transcription-associated amplification.

18. The process of claim 12, wherein the amplification product is a nucleic acid sequence that is the same as or complementary to a sequence of the target nucleic acid, and wherein the probe is hybridized to the amplification product.