Self-contained biological assay apparatus, methods, and applications

• US 9,096,890 B2
• Filed: 02/19/2013
• Issued: 08/04/2015
• 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, Mouchka, Greg, Roswech, Todd, Spizz, Gwendolyn, Yasmin, Rubina
• Primary Examiner(s): Gordon, Brian R
• Application Number:
  US13/770,521
• Publication Number:
  US 20140065598A1
• Time in Patent Office: 896 Days
• Field of Search: 436/43, 436/54, 436/89, 436/180, 436/807, 436/809, 436/811, 436/812, 422500-505, 422507-509, 422/518, 422 62- 67, 422/68.1, 435/287.7
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/141 Preventing contamination, tampering
• CPC Class Codes: B01L 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/088 Channel loops
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0666 Solenoid valves
• CPC Class Codes: B01L 3/022 Capillary pipettes, i.e. having very small bore B01L3/0224 - B01L3/0237 take precedence
• 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 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• 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 1/6806 : Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 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 1/6883 : for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 1/708 : for papilloma
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device
• 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 a pneumatic manifold that operates a microfluidic system having 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, at least one analysis reservoir, and at least one temperature-controlled amplification/reaction reactor attached to 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 reagent reservoir that is in fluid connection with the fluidic transport layer;

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

transporting the fluid test sample to the at least one temperature-controlled amplification/reaction reactor that is in operative communication with the fluidic transport layer;

incubating the fluid test sample in the temperature-controlled amplification/reaction reactor under conditions sufficient to permit the target nucleic acid sequence to be amplified;

transporting the incubated fluid test sample to the at least one analysis reservoir; and

analyzing the amplified target nucleic acid sequence from the incubated test sample,

wherein the test sample is transported from a starting location in the fluidic transport layer to the at least one analysis reservoir separately from the pneumatic manifold and the dispensing system.

2. 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.

3. The automated process of claim 1, wherein the analyzing step further comprises performing a microarray analysis on a microarray in the at least one analysis reservoir in the microfluidic system.

4. The automated process of claim 3, further comprising providing a microarray analysis membrane in the at least one 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 analysis membrane.

5. The automated process of claim 3, further comprising providing a microarray analysis membrane in the at least one analysis reservoir;

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

6. The automated process of claim 3, further comprising providing heat to the at least one analysis reservoir.

7. The automated process of claim 1, wherein analysis of the target nucleic acid sequence indicates the presence of an infectious agent.

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

9. The automated process of claim 1, wherein the analyzing step comprises detecting an interaction between the amplified target nucleic acid sequence and a probe for the target nucleic acid sequence.

10. The automated process of claim 9, wherein the analyzing step comprises determining presence of, or predisposition for:

the disease or disorder of interest,

the infectious agent,

the pathogen,

cancer, or

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

11. The automated process of claim 1, 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.

12. The automated process of claim 11 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.

13. The automated process of claim 1, comprising providing the at least one reagent via a pipette tip, incorporated into the dispensing system.