Nucleic acid amplification and detection using microfluidic diffusion based structures

• US 20010046701A1
• Filed: 05/24/2001
• Published: 11/29/2001
• Est. Priority Date: 05/24/2000
• Status: Abandoned Application

• Current Assignee: Perkinelmer Health Sciences Incorporated (Perkinelmer Incorporated)
• Original Assignee: Perkinelmer Health Sciences Incorporated (Perkinelmer Incorporated)
• Inventors: Weigl, Bernhard H., Schulte, Thomas H.
• Application Number:
  US09/865,093
• Publication Number:
  US 20010046701A1
• Time in Patent Office: Days
• US Class Current: 435/287.2
• CPC Class Codes: B01D 11/00 Solvent extraction
• CPC Class Codes: B01F 2025/913 Vortex flow, i.e. flow spiraling in a tangential direction and moving in an axial direction
• CPC Class Codes: B01F 2025/9171 Parallel flow, i.e. every point of the flow moves in parallel layers where intermixing can occur by diffusion or which do not intermix; Focusing, i.e. compressing parallel layers without intermixing them
• CPC Class Codes: B01F 25/10 Mixing by creating a vortex flow, e.g. by tangential introduction of flow components
• CPC Class Codes: B01F 33/3011 using a sheathing stream of a fluid surrounding a central stream of a different fluid, e.g. for reducing the cross-section of the central stream or to produce droplets from the central stream
• CPC Class Codes: B01F 33/3039 with mixing achieved by diffusion between layers
• CPC Class Codes: B01F 33/834 Mixing in several steps, e.g. successive steps B01F33/81, B01F33/82 and B01F33/85 take precedence
• CPC Class Codes: B01F 35/81 Forming mixtures with changing ratios or gradients
• CPC Class Codes: B01L 13/02 for receptacle or instruments
• CPC Class Codes: B01L 2200/0621 Control of the sequence of chambers filled or emptied
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/0694 Creating chemical gradients in a fluid
• CPC Class Codes: B01L 2300/0809 rectangular shaped
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0638 : membrane valves, flap valves
• CPC Class Codes: B01L 2400/0655 : pinch valves
• 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 3/50273 : 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 3/502776 : specially adapted for focusing or laminating flows
• CPC Class Codes: B01L 3/565 : Seals in general F16L
• 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
• CPC Class Codes: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: F16K 99/0001 : Microvalves microdevices B81B1/00; manufacture or treatment of devices or systems in or on a substrate B81C1/00; microfluidic structures B01L3/5027; micropumps F04B19/006
• CPC Class Codes: F16K 99/0017 : Capillary or surface tension valves, e.g. using electro-wetting or electro-capillarity effects
• CPC Class Codes: F16K 99/0028 : Valves having multiple inlets or outlets
• CPC Class Codes: F16K 99/0057 : the fluid being the circulating fluid itself, e.g. check valves
• CPC Class Codes: G01N 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2035/00247 : Microvalves
• CPC Class Codes: G01N 2035/00514 : Stationary mixing elements
• CPC Class Codes: G01N 35/1097 : characterised by the valves valves in general F16K
• CPC Class Codes: Y10T 137/2076 : Utilizing diverse fluids

Abstract

A device for performing polymerase chain reaction (PCR) amplification and detection using microfluidic diffusion-based structures. Fluid containing DNA to be amplified is cycled repeatedly across hot and cold zones to enhance the multiplication process. The invention is used in conjunction with other devices to perform both single and multiple target detection.

Claims

1. A device for sequentially heating and cooling a fluid, comprising:

a microfluidic channel having a first and a second end;

a fluid specimen located within said channel having a first temperature;

a first region of higher temperature than said first temperature located between said first and second ends of said channel;

a second region of lower temperature than said first temperature located between said first and second ends of said channel

whereby said fluid specimen flows through said channel the temperature of at least a portion of said fluid is sequentially increased and lowered.

2. The device of

claim 1

, wherein said first region comprises a heat strip.

3. The device of

claim 1

, comprising a plurality of first and second regions located between said first and second ends.

4. The device of

claim 1

, wherein said first region is spaced apart from said channel.

7. The device of

claim 1

, wherein said first region of higher temperature comprises approximately 95°

C.

8. The device of

claim 1

, wherein said second region of lower temperature comprises between 45 and 50°

C.

9. The device of

claim 1

, wherein said first region of higher temperature comprises a metal plate.

10. The device of

claim 1

, wherein said first region of higher temperature comprises a radiation heater.

11. The device of

claim 1

, wherein said first region of higher temperature comprises joule heating.

5. A device for sequentially heating and cooling a fluid, comprising:

a microfluidic channel having a first and second end;

and a heating element placed in proximity to said channel,

such that said heating element increases the temperature of portions of said channel at multiple discrete locations.

6. The device of

claim 5

, further comprising a cooling element placed in proximity of said channel such that said cooling element decreases the temperature of portions of said channel at multiple discrete locations.

12. A microfluidic device for performing a polymerase chain reaction to amplify selected nucleic acid sequences, comprising:

a first microfluidic channel containing a selected nucleic acid sequence;

a second microfluidic channel containing substances necessary to perform nucleic acid amplification;

a main microfluidic channel having a sinuous pathway, coupled to said first and second channels such that the contents of said first and second channels establish a laminar flow within said main channel of an initial temperature such that particles may diffuse across the laminar boundary;

a first region of higher temperature than said initial temperature located along said sinuous pathway of said main channel;

a second region of lower temperature than said initial temperature located along said sinuous pathway of said main channel spaced apart from said first region;

whereby the laminar flow within said main channel sequentially passes said first region and said second region such that the temperature of the laminar flow within said main channel is sequentially increased and lowered to amplify the selected nucleic acid sequences.

13. The device of

claim 12

, wherein said substances necessary to perform nucleic amplification include Taq-polymerase.

14. The device of

claim 13

, wherein said substances further include dNTP and two DNA primer sequences.

15. The device of

claim 12

, wherein said main channel is S-shaped.

16. The device of

claim 12

, comprising a plurality of first regions and a plurality of second regions, wherein the locations of said first and second regions are alternated.