Expedited PCR with stirring

Patent metadata

Field	Value
Publication number	US-11155773-B2
Application number	US-201716072932-A
Country	US
Kind code	B2
Filing date	Jan 24, 2017
Priority date	Jan 26, 2016
Publication date	Oct 26, 2021
Grant date	Oct 26, 2021

Abstract

Disclosed are an apparatus and methods for rapid amplification of nucleic acids. More particularly, the present disclosure relates to an apparatus for mixing a reaction solution during amplification of nucleic acids and to methods for amplifying nucleic acids. Also disclosed are methods for lysing cells in a sample and amplifying nucleic acids.

First claim

What is claimed is:

1. An apparatus for conducting amplification of a nucleic acid comprising:
a bit, wherein the bit is coupled to a system configured to apply rotational force to the bit;
a magnetic element that applies a magnetic field to the bit such that the bit is configured to capture one or more paramagnetic particles;
a reaction vessel comprising at least one reagent for amplifying a nucleic acid; and
a measurement system configured to analyze a parameter of the reaction vessel.

2. The apparatus of claim 1 wherein the measurement system comprises a light emitting diode.

3. The apparatus of claim 1 wherein the measurement system comprises a detector for monitoring the amplification.

4. The apparatus of claim 1 wherein the parameter is fluorescence.

5. A method of amplifying a nucleic acid in a sample, the method comprising:
providing a reaction vessel comprising a nucleic acid and at least one reagent for amplifying the nucleic acid;
inserting a portion of a bit into the reaction vessel, wherein the bit is coupled to a system configured to apply rotational force to the bit;
applying a magnetic field to the bit such that the bit is configured to capture one or more paramagnetic particles; and
activating the system to rotate the bit for a sufficient time to amplify the nucleic acid.

6. The method of claim 5 , wherein the bit is rotated at a speed ranging from about 500 rpm to about 7,000 rpm.

7. The method of claim 5 , further comprising monitoring the amplification.

8. The method of claim 7 , wherein the monitoring comprises optical detection.

9. The method of claim 5 , wherein the sample is selected from the group consisting of a saliva sample, a blood sample, a serum sample, a plasma sample and a cheek swab.

10. The method of claim 5 , wherein the sample is selected from the group consisting of a bacterial sample, a viral sample, a fungal sample, a plant sample, and an animal sample.

11. A method for lysing a cell sample and amplifying a nucleic acid, the method comprising:
transferring a sample comprising a cell to a cuvette, wherein the cuvette comprises a lysis buffer;
providing a plurality of paramagnetic particles to the cuvette, wherein the plurality of paramagnetic particles comprises a coating for capturing a nucleic acid in the sample;
inserting a portion of a bit into the lysis buffer, wherein the bit is coupled to a system configured to apply rotational force to the bit and coupled to a magnetic element configured to apply a magnetic field to the bit;
activating the magnetic element to apply the magnet field, resulting in the plurality of paramagnetic particles to contact the bit;
transferring the plurality of magnetic particles to a reaction buffer;
deactivating the magnetic element to release the plurality of paramagnetic particles from the bit; and
activating the system to apply rotational force to the bit for a sufficient time to amplify the nucleic acid.

12. The method of claim 11 , wherein the reaction buffer comprises a reagent for amplification of a nucleic acid.

13. The method of claim 11 , wherein the bit is rotated at a speed ranging from about 500 rpm to about 7,000 rpm.

14. The method of claim 11 , further comprising monitoring the amplification.

15. The method of claim 14 , wherein the monitoring comprises optical detection.

16. The method of claim 11 , wherein the cell sample is selected from the group consisting of a saliva sample, a blood sample, a serum sample, a plasma sample and a cheek swab.

17. The method of claim 11 , wherein the cell sample is selected from the group consisting of a bacterial sample, a viral sample, a fungal sample, a plant sample, and an animal sample.

Assignees

• Nutech Ventures

Inventors

• Viljoen Hendrik

Classifications

• B01L7/5255 Primary

  by moving sample containers · CPC title

• B01L2300/1838

  using fluid heat transfer medium · CPC title

• B01L2400/04

  Moving fluids with specific forces or mechanical means · CPC title

• B01L2400/043

  magnetic forces · CPC title

• B01L2200/0668

  Trapping microscopic beads · CPC title