GENETIC ANALYSIS LOC WITH LOW OLIGONUCLEOTIDE PROBE MASS AND LOW REAGENT VOLUME

• US 20110312765A1
• Filed: 06/01/2011
• Published: 12/22/2011
• Est. Priority Date: 06/17/2010
• Status: Abandoned Application

• Current Assignee: Geneasys Pty Ltd
• Original Assignee: Geneasys Pty Ltd
• Inventors: Silverbrook, Kia, Azimi, Mehdi
• Application Number:
  US13/150,123
• Publication Number:
  US 20110312765A1
• Time in Patent Office: Days
• US Class Current: 506/39
• 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 2300/023 Sending and receiving of information, e.g. using bluetooth
• CPC Class Codes: B01L 2300/024 Storing results with means integrated into the container
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/10 Means to control humidity and/or other gases
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0633 with moving parts
• CPC Class Codes: B01L 2400/0677 phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• 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 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• 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 1/68 involving nucleic acids
• CPC Class Codes: F16K 99/003 Valves for single use only
• CPC Class Codes: F16K 99/0036 operated by temperature variations
• CPC Class Codes: Y02A 90/10 Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation
• CPC Class Codes: Y10T 137/0352 : Controlled by pressure
• CPC Class Codes: Y10T 137/0391 : Affecting flow by the addition of material or energy
• CPC Class Codes: Y10T 137/1044 : With other condition responsive valve control
• CPC Class Codes: Y10T 137/206 : Flow affected by fluid contact, energy field or coanda effect [e.g., pure fluid device or system]
• CPC Class Codes: Y10T 137/2076 : Utilizing diverse fluids
• CPC Class Codes: Y10T 137/2202 : By movable element
• CPC Class Codes: Y10T 436/107497 : Preparation composition [e.g., lysing or precipitation, etc.]
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/145555 : Hetero-N
• CPC Class Codes: Y10T 436/173845 : Amine and quaternary ammonium
• CPC Class Codes: Y10T 436/203332 : Hydroxyl containing
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

A lab-on-a-chip (LOC) device for genetic analysis having a supporting substrate, a microsystems technology (MST) layer supported on the substrate, the MST layer having an inlet for receiving a biological sample containing target nucleic acid sequences, a hybridization section comprising an array of chambers with probes for hybridization with the target nucleic acid sequences to form probe-target hybrids, and, a reagent reservoir containing a reagent necessary for processing the biological sample prior to hybridization, wherein, the hybridization section contains less than 270 picograms of each probe per chamber and the reagent reservoir containing less than 1000,000,000 cubic microns of the reagent.

Claims

1. A lab-on-a-chip (LOC) device for genetic analysis comprising:

a supporting substrate;

a microsystems technology (MST) layer supported on the substrate, the MST layer having an inlet for receiving a biological sample containing target nucleic acid sequences;

a hybridization section comprising an array of chambers with probes for hybridization with the target nucleic acid sequences to form probe-target hybrids; and

,

a reagent reservoir containing a reagent necessary for processing the biological sample prior to hybridization;

wherein,

the hybridization section contains less than 270 picograms of each probe per chamber and the reagent reservoir containing less than 1000,000,000 cubic microns of the reagent.

2. The LOC device according to claim 1 wherein the hybridization section contains less than 60 picograms of each probe per chamber and the reagent reservoir containing less than 300,000,000 cubic microns of the reagent.

3. The LOC device according to claim 1 wherein the hybridization section contains less than 12 picograms of each probe per chamber and the reagent reservoir containing less than 70,000,000 cubic microns of the reagent.

4. The LOC device according to claim 1 wherein the hybridization section contains less than 2.7 picograms of each probe per chamber and the reagent reservoir containing less than 20,000,000 cubic microns of the reagent.

5. The LOC device according to claim 1 further comprising a plurality of reagent reservoirs containing all reagents necessary for performing the diagnostic assay wherein each of the reagent reservoirs have an outlet with a surface tension valve for retaining the reagent in the reservoir with a reagent meniscus until contact with the biological sample removes the reagent meniscus.

6. The LOC device according to claim 5 wherein the MST layer has a flow-path extending from the inlet to the surface tension valves of each of the reagent reservoirs, the flow-path configured to draw the biological sample to the surface tension valves by capillary action.

7. The LOC device according to claim 5 wherein the reagent reservoirs are supported on the MST layer, such that the MST layer is between the reagent reservoirs and the supporting substrate.

8. The LOC device according to claim 7 wherein the reagent reservoirs are defined in a cap, the cap comprising a layer of material with cavities to define the reagent reservoirs.

9. The LOC device according to claim 8 wherein the MST layer has heaters for use in processing the biochemical liquid.

10. The LOC device according to claim 8 wherein at least one of the reagent reservoirs has a plurality of the outlets to increase the reagent flow rate out of the reservoir.

11. The LOC device according to claim 10 wherein the flow-path extends through the MST layer and the cap.

12. The LOC device according to claim 1 wherein the reagent reservoirs each have a vent to atmosphere sized to prevent leakage of the reagent during storage and handling of the LOC device, and allow airflow into the reagent reservoir as the reagent flow out of the outlet.

13. The LOC device according to claim 1 wherein the biochemical liquid is a biological sample and the biochemical processing is a genetic diagnostic assay, the biological sample containing target nucleic acid sequences, the MST layer having probes for hybridization with the target nucleic acid sequences to form probe-target hybrids.

14. The LOC device according to claim 13 further comprising a photosensor wherein the probe-target hybrids each have a fluorophore for emitting a fluorescence signal in response to an excitation light such that the photosensor senses the fluorescence signal to generate an output indicating hybridization of the probes with the target nucleic acid sequence.

15. The LOC device according to claim 13 wherein each hybridization chamber has an optical window for exposing the probes to the excitation light.

16. The LOC device according to claim 15 wherein the MST layer has a polymerase chain reaction (PCR) section for amplifying the oligonucleotides in the sample of biological material.

17. The LOC device according to claim 15 wherein the reagent contained in the reagent reservoir has one or more of:

lysis reagent;

anticoagulant;

polymerase;

dNTP'"'"'s;

primers;

or,

ligase.

18. The LOC device according to claim 17 further comprising CMOS circuitry positioned between the MST layer and the supporting substrate, the CMOS circuitry incorporating the photosensor and bond-pads for communication with an external device.

19. The LOC device according to claim 18 wherein the CMOS circuitry controls activation and deactivation of an external light source configured to generate the excitation light.

20. The LOC device according to claim 19 further comprising an array of hybridization chambers containing different types of the probes, the probes being fluorescence resonance energy transfer (FRET) probes configured for hybridization with different target nucleic acid sequences, and the photo sensor being an array of photodiodes such that each of the hybridization chambers corresponds to a respective one of the photodiodes.