Methods and devices for electromagnetic amplification of nucleic acids

• US 9,044,729 B2
• Filed: 03/15/2013
• Issued: 06/02/2015
• Est. Priority Date: 07/27/2012
• Status: Active Grant

• Current Assignee: International Park Of Creativity
• Original Assignee: International Park Of Creativity
• Inventors: Rengifo, Raul Cuero, Franco Jimenez, Mario Antonio, Gutierrez Calle, Natalia, Sanchez Londono, Mariana
• Primary Examiner(s): MARSCHEL, ARDIN H
• Application Number:
  US13/839,325
• Publication Number:
  US 20140030764A1
• Time in Patent Office: 809 Days
• Field of Search: 435/91.2
• US Class Current: 1/1
• CPC Class Codes: B01J 19/087 employing electric or magnetic energy
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2523/305 Denaturation or renaturation by physical action

Abstract

Methods and devices for amplifying nucleic acids generally involve exposing the nucleic acid to an electromagnetic field, for example a mini-current magnetic field, while performing the steps of PCR. The PCR methods and devices provide numerous advantages over conventional PCR techniques and systems such as reduced reaction times, no heating requirements, and reduced amounts of reagents (e.g., optional use polymerases and primers). Additionally, the PCR methods and devices require significantly shorter reaction times (e.g., less than one hour) compared to conventional PCR techniques and systems (minimum 2 hours). Finally, as shown in the Examples, the PCR methods and devices amplify significantly more DNA compared to conventional PCR techniques and systems. Accordingly, the PCR methods and devices provide a more efficient and cost-effective way to perform PCR when compared to conventional PCR techniques and systems.

Claims

1. A device for amplifying at least one sample of nucleic acid, comprising:

a circuit including at least one conductor through which a current is passed to induce a magnetic field;

a container support adapted to support at least one container of the nucleic acid sample, wherein at least a portion of the container and the sample are positioned within the magnetic field, and wherein the device includes no heating element that the sample is exposed to other than the magnetic-field-inducing circuit; and

a control unit adapted to control the current and the induced magnetic field, wherein the nucleic acid sample is amplified in response to being subjected to the magnetic field, and wherein the magnetic-field-inducing circuit is adapted and operated so that the current is a mini-current and the nucleic-acid amplification is thermal-free.

2. The nucleic-acid amplification device of claim 1, wherein the control unit is adapted to operate the magnetic-field-inducing circuit at a current of about 0.8 A to about 1.0 A to induce a magnetic field of about 0.322 Gauss to about 0.402 Gauss as applied to the sample at a centerline of the container of the sample.

3. The nucleic-acid amplification device of claim 1, wherein the at least one container comprises a plurality of vials, the at least one nucleic acid sample comprises a plurality of nucleic acid samples in the vials, and the device is adapted for batch processing the plurality of nucleic acid samples, and wherein the container support comprises a vial support, the vial support includes an array of holes that each receive a respective one of the vials, the array of holes includes a plurality of rows of the holes, the holes each define a respective center, and the conductor of the magnetic-field-inducing circuit is routed in a position spaced equidistant from each of the hole centers of a closest one of the rows of holes so that the magnetic field is uniformly applied to all of the samples in all of the vials.

4. The nucleic-acid amplification device of claim 3, wherein the conductor of the magnetic-field-inducing circuit is positioned below the vial support and routed centrally between every other one of the rows of the holes.

5. The nucleic-acid amplification device of claim 1, wherein the at least one container comprises at least one tube having a working portion through which the nucleic acid sample flows and the device is adapted for continuous processing, and wherein the container support comprises a tube support, and the conductor of the magnetic-field-inducing circuit and the tube working portion are positioned equidistantly from each other along entire lengths thereof to provide a uniform exposure of the nucleic acid sample in the tube working portion to the magnetic field.

6. The nucleic-acid amplification device of claim 5, wherein the conductor of the magnetic-field-inducing circuit is positioned below the tube support and the tube working portion is positioned above the tube support, with the conductor and tube working portion vertically aligned in conforming arrangements.

7. A method for amplifying a nucleic acid, the method comprising exposing a sample comprising the nucleic acid to a magnetic field using the device of claim 1.

8. The method of claim 7, wherein the sample comprising the nucleic acid further comprises a primer, a polymerase, or a combination thereof.

19. The nucleic-acid amplification device of claim 1, further comprising a cooling system operable to dissipate from the sample any heat generated by the magnetic-field-inducing circuit.

9. A device for amplifying at least one sample of nucleic acid, comprising:

a circuit including at least one conductor through which a mini-current is passed to induce a magnetic field;

at least one tube having a working portion through which the nucleic acid sample flows for continuous processing of the sample;

a tube support adapted to support the tube working portion, and the nucleic acid sample carried therein, within the magnetic field; and

a control unit adapted to control the mini-current and the induced magnetic field, wherein the nucleic acid sample is amplified in response to being subjected to the magnetic field,

wherein the conductor of the magnetic-field-inducing circuit and the tube working portion are positioned to provide a uniform exposure of the nucleic acid sample in the tube working portion to the magnetic field, wherein the device includes no heating element that the sample is exposed to other than the magnetic-field-inducing circuit, and wherein the magnetic-field-inducing circuit is adapted so that the nucleic-acid amplification is thermal-free.

10. The nucleic-acid amplification device of claim 9, wherein the control unit is adapted to operate the magnetic-field-inducing circuit at a mini-current of about 0.8 A to about 1.5 A to induce a magnetic field of about 0.322 Gauss to about 0.61 Gauss as applied to the sample at a centerline of the container of the sample.

11. The nucleic-acid amplification device of claim 9, wherein the control unit is adapted to operate the magnetic-field-inducing circuit at a mini-current of about 0.005 A to about 1.0 A to induce a magnetic field of about 0.001 Gauss to about 0.402 Gauss.

12. The nucleic-acid amplification device of claim 9, wherein the conductor of the magnetic-field-inducing circuit and the tube working portion are positioned equidistantly from each other along entire lengths thereof to provide a uniform exposure of the nucleic acid sample in the tube working portion to the magnetic field.

13. The nucleic-acid amplification device of claim 12, wherein the conductor of the magnetic-field-inducing circuit is positioned below the tube support and the tube working portion is positioned above the tube support, with the conductor and tube working portion vertically aligned in conforming arrangements.

14. The nucleic-acid amplification device of claim 12, wherein the tube working portion is arranged in serpentine configuration and the conductor is arranged in a conforming serpentine configuration so that they are vertically aligned and equidistantly separated along their entire lengths.

15. The nucleic-acid amplification device of claim 9, wherein the control unit include a pumps for flowing the nucleic acid sample through the tube working portion.

16. The nucleic-acid amplification device of claim 9, wherein the tube includes an inlet and an outlet at opposite ends of the tube working portion, the inlet includes a removable closure and is connectable to a sample tank, the outlet includes a removable closure and is connectable to a post-amplification device, and the tube has a diameter and a length selected to provide a predetermined residence time of the sample in the magnetic field for a given sample flow rate;

wherein the control unit includes at least one input, at least one output, and at least one programmed processor;

further comprising a housing for at least the magnetic-field-inducing circuit, the tube working portion, and the tube support; and

further comprising a cooling system including a fan operable by the control unit.

17. A method for amplifying a nucleic acid, the method comprising exposing a sample comprising the nucleic acid to a magnetic field using the device of claim 16.

18. The method of claim 17, wherein the sample comprising the nucleic acid further comprises a primer, a polymerase, or a combination thereof.