System for performing multi-formatted assays

• US 20070243600A1
• Filed: 03/10/2006
• Published: 10/18/2007
• Est. Priority Date: 03/10/2005
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Inventors: Heinz, Robert, Davis, Christopher, Scalese, Robert, Nguyen, Thanh, Li, Florence, Macioszek, Jerzy, Li, Haito, Lair, Gary, Knight, Byron
• Granted Patent: US 8,008,066 B2
• Time in Patent Office: Days
• US Class Current: 435/287.200
• 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 system for processing the contents of multiple reaction receptacles to determine the amount of a target nucleic acid present in one or more of the reaction receptacles or to ascertain the presence or absence of a target nucleic acid in one or more of the reaction receptacles, said system comprising:

(A) at least one temperature-controlled incubator including an incubation chamber for receiving reaction receptacles therein and for exposing the reaction receptacles within said incubation chamber to controlled temperature conditions;

(B) a nucleic acid quantification system constructed and arranged to determine the amount of a target nucleic acid within the reaction receptacles by;

(1) periodically measuring the amount of a signal emitted by the contents of each reaction receptacle; and

(2) quantifying the amount of the target nucleic acid present in each reaction receptacle based upon the periodic measurements; and

(C) a nucleic acid detection system constructed and arranged to ascertain the presence of the target nucleic acid within the reaction receptacle by;

(1) detecting a signal emitted by the contents each reaction receptacle; and

(2) ascertaining the presence or absence of the target nucleic acid in the reaction receptacle based upon whether or not a signal above a threshold level is detected.

2. The system of claim 1, further comprising an automated transport system constructed and arranged to automatically transport reaction receptacles to and from said nucleic acid quantification system and to automatically transport reaction receptacles to said nucleic acid detection system.

3. The system of claim 2, wherein said transport system comprises a transport mechanism rotatable about an axis of rotation.

4. The system of claim 2, wherein:

said nucleic acid quantification system comprises a signal measuring device constructed and arranged to measure the amount of a signal emitted by the contents of a reaction receptacle disposed within said incubation chamber and placed in operative proximity to said signal measuring device; and

said nucleic acid detection system comprises a signal detecting device constructed and arranged to detect a signal emitted by the contents of a reaction receptacle disposed within a detection chamber distinct from said incubation chamber and placed in operative proximity to said signal detecting device.

5. The system of claim 4 further comprising a signal measuring device transport mechanism constructed and arranged to move said signal measuring device into operative proximity with respect to a reaction receptacle disposed within said incubation chamber.

6. The system of claim 5, wherein said signal measuring device comprises a fluorometer constructed and arranged to measure a fluorometric emission from the contents of a reaction receptacle.

7. The system of claim 6, wherein said signal detecting device comprises a luminometer constructed and arranged to detect a chemiluminescent emission from the contents of a reaction receptacle.

8. The system of claim 4, wherein said nucleic acid quantification system and said nucleic acid detection system are contained within a housing.

9. The system of claim 8, wherein said signal measuring device and said signal detecting device are situated on a processing deck within said housing.

10. The system of claim 9, wherein said housing defines a self-contained, stand alone analyzer unit.

11. The system of claim 10, wherein the analyzer unit is movable.

12. The system of claim 1, wherein:

said temperature-controlled incubator includes a reaction receptacle carrier disposed within said incubation chamber and including a plurality of reaction receptacle stations, each of said reaction receptacle stations being constructed and arranged to carry at least one reaction receptacle, and

said nucleic acid quantification system comprises a signal measuring device constructed and arranged to measure the amount of a signal emitted by the contents of a reaction receptacle placed in operative proximity to said signal measuring device, said signal measuring device being positioned with respect to said reaction receptacle carrier such that reaction receptacles carried on said reaction receptacle carrier are successively moved into operative proximity to said signal measuring device as said reaction receptacle carrier moves the reaction receptacles within said incubation chamber, and

wherein said reaction receptacle carrier is controlled so as to move reaction receptacles within said incubation chamber to successively and periodically place reaction receptacles carried thereby in operative proximity to said signal measuring device, and said signal measuring device is controlled so as to make periodic measurements of the amount of signal emitted by the contents of reaction receptacles placed in operative proximity to said signal measuring device.

13. The system of claim 12, said nucleic acid quantification system further comprising a microprocessor adapted to quantify the amount of the target nucleic acid in the reaction receptacles based upon the periodic measurements made by said signal measuring device.

14. The system of claim 12, said nucleic acid quantification system further comprising:

a signal measuring device transport mechanism constructed and arranged to move said signal measuring device with respect to said reaction receptacle carrier,

wherein each reaction receptacle station of said reaction receptacle carrier is constructed and arranged to carry more than one reaction receptacle, said reaction receptacle carrier is controlled so as to successively present said reaction receptacle stations in a signal measuring position with respect to said signal measuring device, and said signal measuring device transport mechanism is controlled so as to move said signal measuring device with respect to said reaction receptacle carrier to successively place said signal measuring device into operative proximity to each of the reaction receptacles carried in said reaction receptacle station moved into the signal measuring position.

15. The system of claim 12, said nucleic acid quantification system comprising more than one signal measuring device, each signal measuring device being constructed and arranged to measure a different signal emitted by the contents of a reaction receptacle.

16. The system of claim 15, said nucleic acid quantification system further comprising:

a signal measuring device transport mechanism constructed and arranged to move said signal measuring devices with respect to said reaction receptacle carrier,

wherein each reaction receptacle station of said reaction receptacle carrier is constructed and arranged to carry more than one reaction receptacle, said reaction receptacle carrier is constructed and arranged to successively present said reaction receptacle stations in signal measuring positions with respect to said signal measuring devices, and said signal measuring device transport mechanism is constructed and arranged to move each of said signal measuring devices with respect to said reaction receptacle carrier to successively place said signal measuring device into operative proximity to each of the reaction receptacles carried in said reaction receptacle station moved into the signal measuring position corresponding to the signal measuring device.

17. The system of claim 12, wherein said reaction receptacle carrier comprises a generally circular carousel mounted within said incubation chamber so as to be rotatable about an axis of rotation, wherein said reaction receptacle stations are positioned around the periphery of said carousel.

18. The system of claim 17, said nucleic acid quantification system further comprising:

a signal measuring device transport mechanism constructed and arranged to move said signal measuring device with respect to said reaction receptacle carrier,

wherein each reaction receptacle station of said reaction receptacle carrier is constructed and arranged to carry more than one reaction receptacle aligned in a radial orientation with respect to the axis of rotation of said carousel, said reaction receptacle carrier is constructed and arranged to successively present said reaction receptacle stations in a signal measuring position with respect to said signal measuring device, and said signal measuring device transport mechanism is constructed and arranged to move said signal measuring device in a generally radial direction with respect to the axis of rotation of said carousel to successively place said signal measuring device into operative proximity to each of the reaction receptacles carried in said reaction receptacle station moved into the signal measuring position.

19. The system of claim 12, wherein said signal measuring device comprises a fluorometer constructed and arranged to measure a fluorometric emission from the contents of a reaction receptacle.

20. The system of claim 19, wherein said fluorometer comprises:

a light emitting element adapted to emit excitation light;

optical elements defining a first optical path, a second optical path, and a third optical path;

a beam splitter; and

a light detecting element, and wherein;

said light emitting element is operatively positioned relative to said first optical path, which is constructed and arranged to transmit at least a portion of the excitation light emitted by said light emitting element,

said beam splitter is operatively positioned relative to said first optical path and said second optical path and is constructed and arranged to receive the excitation light transmitted by said first optical path and to direct at least a portion of the received excitation light having desired excitation spectral components to said second optical path,

said second optical path being positioned relative to a reaction receptacle in operative proximity to the fluorometer so that said second optical path directs the light having desired excitation spectral components toward the reaction receptacle, and said second optical path receives at least a portion of any fluorometric light emitted by the contents of the reaction receptacle and directs the received portion toward said beam splitter,

said beam splitter is operatively positioned relative to said third optical path and is constructed and arranged to receive the fluorometric light transmitted by said second optical path and to direct at least a portion of the received light having desired emission spectral components to said third optical path, and

said light detecting element is operatively positioned relative to said third optical path, which is constructed and arranged to transmit at least a portion of the light having desired emission spectral components toward said light detecting element.

21. The system of claim 20, wherein the optical elements of said first optical path are further constructed and arranged to substantially remove any unwanted spectral components of the light transmitted from the light emitting element.

22. The system of claim 21, wherein the optical elements of said first optical path are further constructed and arranged to condition the light transmitted from said light emitting element so that the light received by said beam splitter from said first optical path is substantially within a prescribed excitation angle range with respect to an optic axis of said first optical path.

23. The system of claim 22, wherein the optical elements of said first optical path comprise:

a light pipe having a receiving end operatively positioned with respect to said light emitting element to receive at least a portion of the excitation light emitted by said light emitting element and to transmit the excitation light received from said receiving end to an opposed end of said light pipe;

a mirror positioned and oriented so as to redirect light emitted by the opposed end of said light pipe;

one or more lenses positioned so as to receive light redirected by said mirror and constructed and arranged to collimate the light received to substantially within the prescribed excitation angle range;

a excitation baffle positioned to receive light collimated by said one or more lenses and constructed and arranged to block substantially all light not within the prescribed excitation angle range; and

an excitation filter positioned to receive light passing through said baffle and constructed and arranged to substantially remove the unwanted spectral components of the excitation light transmitted from said light emitting element.

24. The system of claim 20, wherein the optical elements of said third optical path are constructed and arranged to eliminate at least a portion of any unwanted spectral components of the light transmitted toward the light detecting element.

25. The system of claim 24, wherein the optical elements of said third optical path are further constructed and arranged to condition the light transmitted from said beam splitter so that the light received by said light detecting element from said third optical path is substantially within a prescribed detection angle range with respect to an optic axis of said third optical path.

26. The system of claim 25, wherein the optical elements of said third optical path comprise:

an emission baffle positioned to receive light transmitted from said beam splitter and constructed and arranged to block substantially all light not within the prescribed detection angle range;

an emission filter positioned to receive light passing through said emission baffle and constructed and arranged to substantially remove the unwanted spectral components of the light transmitted toward said light detecting element; and

at least one lens constructed and arranged to focus light passing though said emission filter toward said light detecting element.

27. The system of claim 20, wherein said light emitting element and said light detecting element are connected to a circuit board.

28. The system of claim 20, wherein said beam splitter comprises a dichroic beam splitter.

29. The system of claim 28, wherein said first, second, and third optical paths include first, second, and third optical axes, respectively, and (a) said first optical axis is generally perpendicular to said second optical axis, (b) said second and third optical axes are generally coaxial, and (c) said dichroic beam splitter is oriented at an angle of about 45 degrees with respect to said first, second, and third optical axes.

30. The system of claim 29, wherein said fluorometer comprises:

an excitation component constructed and arranged to direct excitation energy at the reaction receptacle; and

a detection component constructed and arranged to detect the amount of energy emission from the contents of the reaction receptacle.

31. The system of claim 1, said nucleic acid detection system comprising:

a signal detecting device constructed and arranged to detect a signal emitted by the contents of a reaction receptacle placed in operative proximity to said signal detecting device, said signal detecting device being positioned outside said incubation chamber.

32. The system of claim 31, wherein said signal detecting device comprises a luminometer constructed and arranged to detect a chemiluminescent emission from the contents of a reaction receptacle.

33. The system of claim 31, further comprising:

a reaction receptacle transport system including at least one transport mechanism constructed and arranged to transport reaction receptacles to or from said reaction receptacle carrier through a reaction receptacle access opening provided in said incubator and to successively transport reaction receptacles to said signal detecting device following incubation of the reaction receptacles within said incubator.

34. The system of claim 12, wherein said reaction receptacle carrier further comprises magnetic dividers, each being disposed adjacent to one of the reaction receptacles stations, said magnetic dividers each including one or more magnetic elements for exposing a reaction receptacle carried in the adjacent reaction receptacle station to a magnetic field.

35. The system of claim 31, wherein said signal detecting device comprises:

structure defining a transport path along which a plurality of adjacently-arranged reaction receptacles are moved through the device;

a photosensitive device disposed along said transport path and constructed and arranged to detect light emitted from the contents of a reaction receptacle operatively positioned with respect to said photosensitive device; and

a reaction receptacle isolating device positioned adjacent said transport path and constructed and arranged to pivot between;

(1) a first position permitting the plurality of adjacently-arranged reaction receptacles to be moved along said transport path; and

(2) a second position operatively engaging one of said reaction receptacles disposed on said transport path and operatively positioned with respect to said photosensitive device, said reaction receptacle isolating device being constructed and arranged to substantially prevent light from sources other than said operatively positioned reaction receptacle engaged by said reaction receptacle isolating device from being detected by said photosensitive device, said reaction receptacle isolating device being constructed and arranged to pivotally rotate between said first and second positions and including structure at least partially surrounding a reaction receptacle engaged by said reaction receptacle isolating device when said receptacle vessel isolating device is in said second position.

36. The system of claim 35, wherein said signal detecting device further comprises a transport mechanism constructed and arranged to move the plurality of adjacently-arranged reaction receptacles along said transport path in such a manner as to operatively position each of the adjacently-arranged reaction receptacles with respect to said photosensitive device for a time duration sufficient to permit said photosensitive device to detect the amount of light emitted from the contents of the operatively positioned reaction receptacle.

37. The system of claim 35, wherein said signal detecting device further comprises an aperture panel disposed adjacent said transport path and having an aperture formed therein, and wherein said photosensitive device comprises:

a photomultiplier tube adapted to detect light emitted from an object placed before a light-admitting opening at one end of said photomultiplier tube and to generate an electronic signal indicative of light detected by said photomultiplier tube, said photomultiplier tube being positioned on a side of said aperture panel opposite said transport path with said light-admitting opening thereof positioned with respect to said aperture to receive light emitted from the contents of a reaction receptacle disposed on said transport path in front of said aperture; and

a shutter assembly mounted on said aperture panel and constructed and arranged to selectively admit light through said aperture formed in said aperture panel and into said photomultiplier tube by movement of said shutter assembly between an open position allowing light to pass through said aperture and a closed position preventing light from passing through said aperture, said shutter assembly comprising;

a shutter constructed and arranged for rotational movement between open and closed positions corresponding to open and closed positions, respectively, of said shutter assembly, whereby said shutter blocks said aperture when said shutter is in said closed position and does not block said aperture when said shutter is in said open position; and

a motor operatively coupled to said shutter for effecting powered rotation of said shutter between said open and closed positions.

38. The system of claim 1 further comprising a separation device constructed and arranged isolate a solid support material for binding a target nucleic acid present in a reaction receptacle.

39. The system of claim 38, wherein said separation device includes a fluid aspirator mechanism constructed and arranged to aspirate fluid material from the reaction receptacle.

40. The system of claim 39, wherein said separation device further comprises:

a fluid dispense mechanism constructed and arranged to provide a wash solution to the reaction receptacle after removing the fluid sample from the reaction receptacle; and

a mixing device constructed and arranged to agitate the reaction receptacle to resuspend the solid support material after the wash solution has been provided by the fluid dispense mechanism.

41. The system of claim 38, wherein the separation device comprises magnetic elements for subjecting a fluid material to a magnetic field.

42. The system of claim 1, wherein each reaction receptacle is one of a plurality of integrally formed reaction receptacles.