Apparatus and method for integrated sample preparation, reaction and detection

• US 9,931,636 B2
• Filed: 12/18/2015
• Issued: 04/03/2018
• Est. Priority Date: 02/23/2010
• Status: Active Grant

• Current Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Original Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Inventors: Ching, Jesus, Lee, Phillip You Fai, Richardson, Bruce
• Primary Examiner(s): Hyun, Paul S
• Application Number:
  US14/974,382
• Publication Number:
  US 20160101421A1
• Time in Patent Office: 837 Days
• CPC Class Codes: B01F 31/65 the materials to be mixed being directly submitted to a pulsating movement, e.g. by means of an oscillating piston or air column
• CPC Class Codes: B01F 35/712 for feeding fluids
• CPC Class Codes: B01F 35/715 Feeding the components in several steps, e.g. successive steps
• CPC Class Codes: B01F 35/7174 using pistons, plungers or syringes
• CPC Class Codes: B01L 2200/026 Fluid interfacing between devices or objects, e.g. connectors, inlet details
• CPC Class Codes: B01L 2200/028 Modular arrangements
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• 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 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/042 Caps; Plugs
• CPC Class Codes: B01L 2300/0672 Integrated piercing tool
• 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 2400/0478 pistons
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 2400/0622 distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 2400/0644 rotary valves
• CPC Class Codes: B01L 2400/0683 mechanically breaking a wall or membrane within a channel or chamber
• CPC Class Codes: B01L 3/0265 using valves to interrupt or meter fluid flow, e.g. using solenoids or metering valves
• CPC Class Codes: B01L 3/502 : with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• 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: B01L 7/525 : with physical movement of samples between temperature zones

Abstract

An apparatus includes a housing, a reaction vial and a transfer mechanism. The housing defines a first flow path and a second flow path. The housing has transfer port defining an opening in fluid communication with the second flow path and a volume outside of the housing. The transfer port includes a flow control member to limit flow through the opening. The reaction vial is coupled to the housing and defines a reaction volume, which is in fluid communication with the transfer port via the second flow path. The transfer mechanism is configured to transfer a sample from an isolation chamber of an isolation module to the reaction chamber via at least the first flow path when the transfer mechanism is actuated. The transfer mechanism configured to produce a vacuum in the reaction vial to produce a flow of a sample from the isolation chamber to the reaction volume.

Claims

1. An apparatus, comprising:

a housing defining a linear flow path;

a reaction vial coupled to the housing, the reaction vial defining a reaction volume, the reaction volume in fluid communication with the linear flow path, wherein the reaction vial comprises a lyophilized substance comprising PCR reagents;

a transfer mechanism configured to transfer a sample from the reaction vial into the linear flow path when the transfer mechanism is actuated; and

a plurality of movable members movably coupled to the housing, the plurality of movable members configured to separate the linear flow path into a plurality of PCR volumes, each PCR volume from the plurality of PCR volumes being fluidically isolated from an adjacent PCR volume from the plurality of PCR volumes.

2. The apparatus of claim 1, wherein each movable member from the plurality of movable members is configured to move between a first position and a second position, the plurality of movable members configured to separate the linear flow path into a plurality of PCR volumes when the plurality of movable members is in the second position.

3. The apparatus of claim 1, wherein:

the linear flow path is a first flow path;

the transfer mechanism is a first transfer mechanism; and

the housing defines a second flow path, the housing configured to be coupled to an isolation module such that the sample can be conveyed from an isolation chamber of the isolation module to the reaction volume via the second flow path when a second transfer mechanism is actuated.

4. The apparatus of claim 1, wherein a first movable member from the plurality of movable members is configured to move between a first position and a second position independently from the movement of a second movable member from the plurality of movable members.

5. The apparatus of claim 1 wherein each PCR volume is approximately 20 microliters.

6. The apparatus of claim 1 wherein each PCR volume is between approximately 500 nL and approximately 10 microliters.

7. The apparatus of claim 1 wherein each PCR volume is between approximately 500 nL and approximately 5 microliters.

8. A method, comprising:

using an apparatus comprising;

a housing defining a linear flow path;

a reaction vial coupled to the housing, the reaction vial defining a reaction volume, the reaction volume in fluid communication with the linear flow path, wherein the reaction vial comprises a lyophilized substance comprising PCR reagents;

a transfer mechanism configured to transfer a sample from the reaction vial into the linear flow path when the transfer mechanism is actuated; and

a plurality of movable members movably coupled to the housing, the plurality of movable members configured to separate the linear flow path into a plurality of PCR volumes, each PCR volume from the plurality of PCR volumes being fluidically isolated from an adjacent PCR volume from the plurality of PCR volumes;

conveying the sample from the reaction volume into the linear flow path defined by the housing, the sample including a plurality of target nucleic acid molecules;

moving the plurality of movable members to divide the linear flow path into the plurality of PCR volumes such that each PCR volume from the plurality of PCR volumes includes no more than one target nucleic acid molecule from the plurality of target nucleic acid molecules; and

activating a heating element to thermally cycle the contents of each PCR volume from the plurality of PCR volumes.

9. The method of claim 8, wherein the moving includes moving a first movable member from the plurality of movable members at a different time than moving a second movable member from the plurality of movable members.

10. The method of claim 8, further comprising:

heating the sample within the reaction volume before the conveying.

11. The method of claim 8 wherein each PCR volume is approximately 20 microliters.

12. The method of claim 8 wherein each PCR volume is between approximately 500 nL and approximately 10 microliters.

13. The method of claim 8 wherein each PCR volume is between approximately 500 nL and approximately 5 microliters.