Method for continuous mode processing of the contents of multiple reaction receptacles in a real-time amplification assay

• US 8,349,564 B2
• Filed: 11/04/2010
• Issued: 01/08/2013
• Est. Priority Date: 03/10/2005
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Inventors: Macioszek, Jerzy A., Davis, Christopher B., Lair, Gary D., Li, Haitao, Li, Florence F., Knight, Byron J., Scalese, Robert F., Heinz, Robert E., Nguyen, Thanh N.
• Primary Examiner(s): BERTAGNA, ANGELA MARIE
• Application Number:
  US12/939,420
• Publication Number:
  US 20110053169A1
• Time in Patent Office: 796 Days
• Field of Search: None
• US Class Current: 435/6.12
• 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. An automated method for processing the contents of multiple reaction receptacles to determine the presence or amount of a target nucleic acid in each of the reaction receptacles, the method comprising the steps of:

a) isolating and purifying target nucleic acid that may be present in a sample material contained within each of the reaction receptacles by;

i) exposing each reaction receptacle to a magnetic field, thereby isolating magnetically responsive solid supports within each reaction receptacle, the solid supports being capable of directly or indirectly immobilizing the target nucleic acid thereon;

ii) during step i), removing at least a portion of the sample material in each reaction receptacle from the isolated solid supports; and

iii) after step ii), removing the magnetic field and washing the solid supports in each reaction receptacle with a wash solution;

b) moving the reaction receptacles to a temperature-controlled chamber of an incubator, each reaction receptacle being provided with reagents sufficient to amplify and detect the presence of a target sequence contained within the target nucleic acid under the temperature conditions of the incubator, and effecting amplification of the target nucleic acid;

c) exposing each reaction receptacle to a magnetic field within the incubator, thereby aggregating at least a portion of the solid supports within each reaction receptacle, wherein the incubator includes a reaction receptacle carrier that is rotatable about an axis of rotation and is configured to receive and hold reaction receptacles moved to the incubator, and wherein the receptacle carrier comprises;

i) a plurality of receptacle carrier stations, each station being configured to receive and hold at least one reaction receptacle; and

ii) a magnetic divider disposed between each of a pair of adjacent carrier stations and including one or more magnets configured to expose each reaction receptacle carried in one of the pair of adjacent carrier stations to a magnetic field in accordance with step c); and

d) during step c), measuring the amount of a signal associated with an amplification product free in solution containing the target sequence or its complement in each reaction receptacle provided to the incubator in accordance with step b).

2. The method of claim 1, wherein the carrier comprises a circular carousel that is oriented horizontally and is rotatable about a vertical axis of rotation with the receptacle carrier stations arranged around the circumference of the carrousel, and wherein the magnetic dividers are angularly spaced about the axis of rotation.

3. The method of claim 2, wherein each magnetic divider comprises a divider wall that is oriented radially with respect to the axis of rotation.

4. The method of claim 1, wherein each magnet comprises neodymium-iron-boron.