Microfluidic system for nucleic acid analysis

Patent metadata

Field	Value
Publication number	US-9657341-B2
Application number	US-201314074590-A
Country	US
Kind code	B2
Filing date	Nov 7, 2013
Priority date	Jan 24, 2013
Publication date	May 23, 2017
Grant date	May 23, 2017

Abstract

A microfluidic system for analyzing nucleic acid, the microfluidic system including a reagent supply device including a sample chamber into which a sample can be injected, one or more reagent chambers for containing one or more reagents for extracting nucleic acid from the sample, and a waste chamber in which the used reagent can be discarded; a binding-lysis chamber in which cells are captured from the sample and lysed to form a cell lysate containing nucleic acid; plurality of particles for cell binding disposed in the binding-lysis chamber; a plurality of rehydration chambers into which the cell lysate formed in the binding-lysis chamber can be distributed and mixed with a nucleic acid amplification reagent to form an amplification reaction mixture; a plurality of amplification chambers in which a nucleic acid amplification reaction is performed on the amplification reaction mixture introduced from the plurality of rehydration chambers; and a flow channel system including an outlet and a plurality of inlets connected to the reagent supply device and forming an integrated fluid flow between the binding-lysis chamber, the rehydration chambers, and the amplification chambers.

First claim

What is claimed is:

1. A microfluidic system for analyzing nucleic acid, the microfluidic system comprising:
a reagent supply device including a sample chamber into which a sample can be injected, one or more reagent chambers for containing one or more reagents for extracting nucleic acid from the sample, and a waste chamber in which the used reagent can be discarded;
a binding-lysis chamber in which cells are captured from the sample and lysed to form a cell lysate containing nucleic acid;
a plurality of particles for cell binding disposed in the binding-lysis chamber;
a plurality of rehydration chambers into which the cell lysate formed in the binding-lysis chamber can be distributed and mixed with a nucleic acid amplification reagent to form an amplification reaction mixture;
a plurality of amplification chambers in which a nucleic acid amplification reaction is performed on the amplification reaction mixture introduced from the plurality of rehydration chambers; and
a flow channel system including an outlet and a plurality of inlets connected to the reagent supply device and forming an integrated fluid flow between the binding-lysis chamber, the rehydration chambers, and the amplification chambers,
and further comprising a fluid flow part comprising:
a top surface comprising the plurality of inlets and the outlet, which are connected to the reagent supply device;
a first through hole corresponding to the binding-lysis chamber;
a plurality of second through holes corresponding to the plurality of rehydration chambers; and
a bottom surface comprising a recessed groove pattern comprising a plurality of recessed grooves corresponding to the plurality of nucleic acid amplification chambers;

a membrane part comprising an elastic membrane bonded to the bottom surface of the fluid flow part to form bottom surfaces of the binding-lysis chamber and the plurality of rehydration chambers;
a pneumatic part bonded to a bottom surface of the membrane part and having a plurality of ports for applying pneumatic pressure to one or more predetermined positions of the membrane part; and
a guide part for installing the reagent supply device, wherein the guide part is disposed on an upper portion of the fluid flow part and configured to align the plurality of inlets and the outlet of the fluid flow part with the plurality of chambers of the reagent supply part.

2. The microfluidic system of claim 1 , wherein the plurality of reagent chambers comprises a lysis buffer chamber in which a lysis buffer can be injected, and a washing buffer chamber in which a washing buffer can be injected.

3. The microfluidic system of claim 2 , wherein a destruction pattern, which is to be broken by external impact, is formed on each bottom surface of the sample chamber, the lysis buffer chamber, the washing buffer chamber, and the waste chamber.

4. The microfluidic system of claim 3 , wherein the outlet and the plurality of inlets have a shape of a needle for applying an impact onto the destruction pattern.

5. The microfluidic system of claim 2 , further comprising one or more metering chambers for quantifying an amount of the lysis buffer supplied from the lysis buffer chamber of the reagent supply device.

6. The microfluidic system of claim 1 , further comprising one or more bubble trap chambers for removing bubbles generated during cell lysis in the binding-lysis chamber.

7. The microfluidic system of claim 1 , wherein each of the plurality of rehydration chambers comprises two separated subchambers, and the nucleic acid amplification reagent is divided and disposed in the two subchambers.

8. The microfluidic system of claim 7 , wherein, in each of the plurality of rehydration chambers, a sample including nucleic acid is disposed in one subchamber and a reagent including an enzyme is disposed in the other subchamber.

9. The microfluidic system of claim 8 , wherein the sample including nucleic acid comprises one or more of a probe and a primer.

10. The microfluidic system of claim 7 , wherein the nucleic acid amplification reagent is in a freeze-dried form.

11. The microfluidic system of claim 7 , wherein a side of the subchamber has a curved shape and comprises a flow path therethrough with a width that is smallest at a center portion of the subchamber.

12. The microfluidic system of claim 1 , further comprising a plurality of metering chambers for quantifying an amount of the cell lysate formed in the binding-lysis chamber and distributing the cell lysate into the plurality of rehydration chambers.

13. The microfluidic system of claim 1 further comprising:
a binding-lysis chamber cover member that covers the first through hole at the top surface of the fluid flow part, wherein the cover member, the first through hole, and the membrane part define the binding lysis-chamber;
one or more rehydration chamber cover members that cover the plurality of second through holes at the top surface of the fluid flow part, wherein each of the second through holes together with the cover member and membrane part define a rehydration chamber;
a PCR film positioned on the bottom surface of the fluid flow part to cover the recessed groove pattern, wherein each of the recessed grooves together with the PCR film define a nucleic acid amplification chamber.

14. The microfluidic system of claim 1 , wherein the bottom surface of the fluid flow part further comprises a microchannel providing the flow channel system, and a microvalve for preventing flow of a fluid passing along the microchannel when pneumatic pressure is applied from the pneumatic part.

15. The microfluidic system of claim 1 , wherein a plurality of particles for cell binding is disposed in the first through hole.

16. The microfluidic system of claim 13 , wherein the top surface of the fluid flow part comprises a bridge pattern having a shape recessed in the top surface of the fluid flow part that forms a path from the rehydration chamber to the nucleic acid amplification chamber by which the amplification reaction mixture can be transferred from the rehydration chamber to the nucleic amplification chamber.

17. The microfluidic system of claim 1 , wherein the bottom surface of the fluid flow part further comprises a recess pattern,
the recess pattern providing one or more metering chambers for quantifying an amount of the lysis buffer supplied from the lysis buffer chamber of the reagent supply device on the bottom surface of the fluid flow part, one or more bubble trap chambers for removing bubbles generated during cell lysis in the binding-lysis chamber on the bottom surface of the fluid flow part, and a plurality of metering chambers for quantifying an amount of the cell lysate formed in the binding-lysis chamber and distributing the cell lysate into the plurality of rehydration chambers.

18. The microfluidic system of claim 1 , wherein the fluid flow part is formed of a transparent polymer material.

19. The microfluidic system of claim 1 , wherein the fluid flow part is formed of any one of polycarbonate (PC), polymethyl methacrylate (PMMA), polystyrene (PS), cyclic olefin copolymer (COC), polydimethylsiloxane (PDMS), and silicone.

20. The microfluidic system of claim 1 , wherein the membrane part is formed of PDMS or silicone.

21. The microfluidic system of claim 1 , wherein the pneumatic part is formed of a transparent polymer material.

22. A microfluidic system for analyzing nucleic acid, the microfluidic system comprising:
a reagent supply device

Assignees

• Samsung Electronics Co Ltd

Inventors

• Park Chin-Sung
• Namkoong Kak
• Shim Joon-Sub
• Chung Won-Seok
• Kim Kyung-Ho
• Kim Joon-Ho
• Jung Won-Jong

Classifications

• B01L3/5027 Primary

  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 · CPC title

• B01L2300/0867

  Multiple inlets and one sample wells, e.g. mixing, dilution · CPC title

• B01L2200/025

  Align devices or objects to ensure defined positions relative to each other · CPC title

• B01L2200/028

  Modular arrangements · CPC title

• B01L7/525

  with physical movement of samples between temperature zones · CPC title