Self-contained biological assay apparatus, methods, and applications

• US 9,102,979 B2
• Filed: 01/30/2015
• Issued: 08/11/2015
• Est. Priority Date: 02/23/2010
• Status: Active Grant

• Current Assignee: Rheonix, Inc.
• Original Assignee: Rheonix, Inc.
• Inventors: Zhou, Peng, Young, Lincoln C., Thomas, Benjamin, Chen, Zongyuan, Roswech, Todd, Spizz, Gwendolyn, Yasmin, Rubina, Mouchka, Greg
• Primary Examiner(s): Gordon, Brian R
• Application Number:
  US14/609,697
• Publication Number:
  US 20150141291A1
• Time in Patent Office: 193 Days
• Field of Search: 436/43, 436/54, 436/89, 436/164, 436/166, 436/172, 436/180, 436/807, 436/809, 436/811, 436/812, 422500-505, 422507-509, 422/518, 422 62- 67, 422/68.1, 422/82.05, 422/82.08, 435/287.7
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/14 Process control and prevention of errors
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0655 pinch valves
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/523 with means for closing or opening
• CPC Class Codes: B01L 3/527 for a plurality of reagents
• 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 1/6809 Methods for determination or identification of nucleic acids involving differential detection
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6846 : Common amplification features
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6876 : Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 1/708 : for papilloma
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device
• CPC Class Codes: C12Q 2600/106 : Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/156 : Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/158 : Expression markers
• CPC Class Codes: G01N 2035/00346 : Heating or cooling arrangements
• CPC Class Codes: G01N 35/10 : Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices G01N35/0099 takes precedence
• CPC Class Codes: G01N 35/1002 : Reagent dispensers

Abstract

A self-contained, fully automated, biological assay-performing apparatus includes a housing; a dispensing platform including a controllably-movable reagent dispensing system, disposed in the housing; a reagent supply component disposed in the housing; a pneumatic manifold removably disposed in the housing in a space shared by the dispensing platform, removably coupled to a fluidic transport layer and a plurality of reservoirs, wherein the fluidic transport layer, the reservoirs, and a test sample to be introduced therein are disposed in the housing in the space separate from the dispensing platform; a pneumatic supply system removably coupled to the pneumatic manifold in the housing in a space separate from the dispensing platform; and a control system coupled to at least one of the dispensing platform and the pneumatic supply system, disposed in the housing.

Claims

1. An automated process for isolating, amplifying and analyzing a target nucleic acid sequence that may be present in a fluid test sample, comprising:

providing a dispensing system;

providing an optical unit comprising a light source and a detector;

providing a pneumatic manifold that operates a microfluidic system having at least one amplification/reaction reactor, a fluidic transport layer and at least one fluidic channel disposed in the fluidic transport layer;

at least one sample input reservoir, at least one reagent reservoir, and at least one analysis reservoir in fluid connection with the fluidic transport layer;

introducing the fluid test sample into the at least one fluidic channel;

providing at least one reagent to the at least one fluidic channel from at least one reservoir that is in fluid connection with the fluidic transport layer and the fluidic channel;

combining the fluid test sample and the at least one reagent in the fluidic transport layer, the at least one fluidic channel, the at least one sample input reservoir or amplification/reaction reactor;

transporting the fluid test sample to the amplification/reaction reactor that is in operative communication with the fluidic transport layer;

incubating the fluid test sample in the amplification/reaction reactor under conditions sufficient to permit the target nucleic acid sequence to be amplified in the presence of a fluorescent reporter;

illuminating, by the light source, at least a portion of the amplification/reaction reactor; and

measuring, by the detector, light emission from the fluorescent reporter in response to illumination by the light source, wherein said measuring is performed in real time during amplification of the target nucleic acid sequence.

2. The automated process of claim 1, further comprising:

transporting the amplified target nucleic acid sequence from the amplification/reaction reactor to the at least one analysis reservoir; and

analyzing the amplified target nucleic acid sequence from the test sample in the at least one analysis reservoir separately from any other samples and separately from the pneumatic manifold and the dispensing system.

3. The automated process of claim 1, wherein the transporting and combining steps are accomplished by pumping the fluid test sample and the volume of the at least one reagent through the fluidic transport layer via at least one multi-valve diaphragm pump that is operated by the pneumatic manifold.

4. The automated process of claim 2, wherein the analyzing step further comprises performing a microarray analysis in a microarray analysis reservoir in the microfluidic system.

5. The automated process of claim 4, further comprising providing a microarray analysis membrane in the microarray analysis reservoir;

flowing a fluid over a top surface of the microarray analysis membrane; and

removing the fluid substantially through a fluid exit route along a periphery of the microarray analysis membrane.

6. The automated process of claim 4, further comprising providing a microarray analysis membrane in the microarray analysis reservoir;

and, flowing a fluid alternatively back and forth over a top surface of the microarray analysis membrane.

7. The automated process of claim 4, further comprising providing heat to the microarray analysis membrane.

8. The automated process of claim 1, wherein results from measuring the target nucleic acid sequence are indicative of a disease or disorder of interest, an infectious agent, a pathogen, a predisposition for cancer, or a predisposition for sensitivity to a drag, pharmaceutical composition, chemical or compound of interest.

9. The automated process of claim 1, wherein the target nucleic acid sequence comprises a SNP.

10. The automated process of claim 8, wherein the disease or disorder is HPV or sepsis.

11. The automated process of claim 8, wherein the target nucleic acid sequence indicates a predisposition for warfarin sensitivity or predisposition for anticoagulation in response to warfarin treatment.

12. The automated process of claim 2, wherein the analyzing step comprises detecting an interaction between the amplified target nucleic acid sequence and a probe for the target nucleic acid sequence.

13. The automated process of claim 12, wherein the analyzing step comprises determining presence of, or predisposition for:

a disease or disorder of interest,

an infectious agent,

a pathogen,

cancer, or

sensitivity to the drug, pharmaceutical composition, chemical or compound of interest.

14. The automated process of claim 2, wherein the analyzing step comprises determining an amount or level of the amplified target nucleic acid sequence and wherein the method further comprises comparing the amount or level with a preselected amount or level of the target nucleic acid sequence.

15. The automated process of claim 14, wherein a difference between the amount or level and the preselected amount or level is indicative of presence of, or predisposition for:

a disease or disorder of interest,

an infectious agent,

a pathogen,

cancer, or

sensitivity to a drug, pharmaceutical composition, chemical or compound of interest.