Genetic test module with low oligonucleotide probe mass and reagent volumes

• US 8,383,064 B2
• Filed: 06/01/2011
• Issued: 02/26/2013
• Est. Priority Date: 06/17/2010
• Status: Expired due to Fees

• Current Assignee: Geneasys Pty Ltd
• Original Assignee: Silverbrook Research Pty Limited
• Inventors: Azimi, Mehdi, Silverbrook, Kia
• Primary Examiner(s): Bullock, In Suk
• Assistant Examiner(s): Kingan, Timothy G
• Application Number:
  US13/150,135
• Publication Number:
  US 20110312589A1
• Time in Patent Office: 636 Days
• Field of Search: 422/68.1, 422/402, 422/417, 422/419, 422/503, 422/527, 422/537, 422/552, 435/287.2, 436/94, 436/96, 436/111, 436/131, 436/527, 506/16
• US Class Current: 422/503
• 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 test module for performing a genetic diagnostic assay, the test module having an outer casing dimensioned for hand-held portability, the outer casing having a receptacle for a biological sample containing target nucleic acid sequences, an array of chambers containing probes for hybridization with the target nucleic acid sequences to form probe-target hybrids, a flow-path extending from the inlet to the probes, and, a reagent reservoir containing a reagent for addition to the sample in the flow-path upstream of the probes, wherein, each of the chambers contains less than 270 picograms of probe and the reagent reservoir has a volume less than 1000,000,000 cubic microns.

Claims

1. A test module for performing a genetic diagnostic assay, the test module comprising:

an outer casing dimensioned for hand-held portability, the outer casing having a receptacle for a biological sample containing target nucleic acid sequences;

an array of chambers containing probes for hybridization with the target nucleic acid sequences to form probe-target hybrids, each of the chambers containing less than 270 picograms of probe;

a flow-path extending from the receptacle to the probes; and

,

a reagent reservoir containing a reagent for addition to the sample in the flow-path upstream of the probes, the reagent reservoir having a volume less than 1000,000,000 cubic microns;

wherein,

the reagent reservoir has 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.

2. A test module according to claim 1 wherein each of the chambers contains less than 60 picograms of probe and the reagent reservoir has a volume less than 300,000,000 cubic microns.

3. A test module according to claim 1 wherein each of the chambers contains less than 12 picograms of probe and the reagent reservoir has a volume less than 70,000,000 cubic microns.

4. A test module according to claim 1 wherein each of the chambers contains less than 2.7 picograms of probe and the reagent reservoir has a volume less than 20,000,000 cubic microns.

5. A test module according to claim 1 further comprising a flow-path extending from the receptacle to the surface tension valve, the flow-path being configured to draw the biological sample to the surface tension valve by capillary action.

6. A test module according to claim 5 further comprising a microfluidic device supported in the casing, the microfluidic device having a supporting substrate and a MST layer formed on the substrate, wherein the reagent reservoir is supported on the MST layer, such that the MST layer is between the reagent reservoir and the supporting substrate.

7. A test module according to claim 6 wherein the reagent reservoir is defined in a cap, the cap comprising a layer of material with a cavity to define the reagent reservoir.

8. A test module according to claim 7 wherein the cap defines a plurality of the reagent reservoirs for containing all the reagents required for the genetic analysis to be performed.

9. A test module 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.

10. A test module according to claim 9 wherein the flow-path extends through the MST layer and the cap.

11. A test module according to claim 1 wherein the reagent reservoir has a vent to atmosphere sized to prevent leakage of the reagent during storage and handling of the microfluidic device, and allow airflow into the reagent reservoir as the reagent flow out of the outlet.

12. A test module according to claim 6 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.

13. A test module according to claim 12 further comprising an LED for generating the excitation light.

14. A test module according to claim 12 wherein each of the chambers is a hybridization chamber, the hybridization chamber having an optical window for exposing the probes to the excitation light.

15. A test module according to claim 14 wherein the MST layer has a polymerase chain reaction (PCR) section for amplifying the oligonucleotides in the sample of biological material.

16. A test module according to claim 14 wherein the reagent contained in the reagent reservoir has one or more of:

lysis reagent;

anticoagulant;

polymerase;

dNTP'"'"'s;

primers;

or,

ligase.

17. A test module according to claim 16 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.

18. A test module according to claim 17 wherein the CMOS circuitry controls activation and deactivation of an external light source configured to generate the excitation light.

19. A test module according to claim 18 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 photosensor being an array of photodiodes such that each of the hybridization chambers corresponds to a respective one of the photodiodes.