Device and method for multiple analyte detection

• US 7,687,280 B2
• Filed: 05/29/2007
• Issued: 03/30/2010
• Est. Priority Date: 04/03/1996
• Status: Expired due to Term

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Woudenberg, Timothy, Kowallis, Reid B., Albin, Michael, Winn-Deen, Emily S., Ragusa, Robert P., Raysberg, Yefim
• Primary Examiner(s): Nguyen; Bao-Thuy L
• Assistant Examiner(s): DIRAMIO, JACQUELINE A
• Application Number:
  US11/807,536
• Publication Number:
  US 20080108068A1
• Time in Patent Office: 1,036 Days
• Field of Search: 435/4, 435/6, 435/7.1, 435/287.2, 435/288.4, 435/288.5, 435/288.7, 436/501, 436/518, 436/164, 436/166, 436/807, 433/50, 433/61, 433/68.1, 422/50, 422/55, 422/57, 422/58, 422/68.1, 422/102, 422/82.05, 422/82.08
• US Class Current: 436/501
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00599 Solution-phase processes
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0642 Filling fluids into wells by specific techniques
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/049 vacuum
• 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 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• 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 1/08 using multifield media
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2565/631 being a biochannel or pore
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 50/08 Liquid phase synthesis, i.e. wherein all library building blocks are in liquid phase or in solution during library creation; Particular methods of cleavage from the liquid support
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 2035/00148 Test cards, e.g. Biomerieux or McDonnel multiwell test cards
• CPC Class Codes: G01N 21/6452 : Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: Y10S 436/805 : Optical property
• CPC Class Codes: Y10S 436/807 : Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/808 : Automated or kit
• CPC Class Codes: Y10S 436/809 : Multifield plates or multicontainer arrays
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/25 : including sample preparation

Abstract

The invention is directed to a method and device for simultaneously testing a sample for the presence, absence, and/or amounts of one or more of a plurality of selected analytes. The invention includes, in one aspect, a device for detecting or quantitating a plurality of different analytes in a liquid sample. Each chamber may include an analyte-specific reagent effective to react with a selected analyte that may be present in the sample, and detection means for detecting the signal. Also disclosed are methods utilizing the device.

Claims

1. A method of detecting or quantitating a target nucleic acid sequence in a liquid sample, comprising:

introducing a liquid sample comprising a target nucleic acid sequence through a channel in a substrate and into a chamber in the substrate which is in fluid communication with the channel, the chamber being pre-loaded with detection reagents before introduction of the liquid sample, the detection reagents comprising a first unlabeled oligonucleotide, a second unlabeled oligonucleotide, and a fluorescently-labeled oligonucleotide;

mixing the liquid sample with the detection reagents in the chamber such that the first unlabeled oligonucleotide, the second unlabeled oligonucleotide, and the fluorescently-labeled oligonucleotide are not immobilized in the chamber;

reacting the target nucleic acid sequence with both the first unlabeled oligonucleotide and the second unlabeled oligonucleotide, in the chamber, under conditions effective to selectively bind each of the first unlabeled oligonucleotide and the second unlabeled oligonucleotide to the target nucleic acid sequence, and to produce a first reaction product;

producing fluorescence in the chamber by a reaction in the chamber involving the first reaction product and the non-immobilized fluorescently-labeled oligonucleotide; and

detecting the fluorescence produced in the chamber.

2. The method of claim 1, wherein the target nucleic acid sequence comprises a single-stranded nucleic acid sequence.

3. The method of claim 1, wherein the first unlabeled oligonucleotide and the second unlabeled oligonucleotide, each bind to a different region of the target nucleic acid sequence.

4. The method of claim 1, further comprising reacting to bind the fluorescently-labeled oligonucleotide to the target nucleic acid sequence such that the fluorescently labeled oligonucleotide emits a detectable fluorescent signal upon excitation thereof.

5. The method of claim 1, wherein the reacting comprises a polymerase chain reaction.

6. The method of claim 1, wherein the chamber comprises a plurality of chambers.

7. The method of claim 1, wherein the chamber comprises a plurality of chambers connected to a channel.

8. The method of claim 1, wherein the introducing a liquid sample comprises subjecting the liquid sample to a vacuum.

9. The method of claim 1, wherein the first unlabeled oligonucleotide and the second unlabeled oligonucleotide comprise a primer pair for polymerase chain reaction amplification.

10. The method of claim 1, wherein the fluorescently-labeled oligonucleotide is a sequence-selective oligonucleotide.

11. The method of claim 10, wherein the sequence-selective oligonucleotide comprises a fluorescer-quencher pair.

12. The method of claim 1, wherein the first oligonucleotide and second oligonucleotide bind selectively to adjacent, contiguous regions of the target nucleic acid.

13. The method of claim 1, wherein the detection reagents are dried detection reagents.