Diagnostic and sample preparation devices and methods

• US 10,301,666 B2
• Filed: 01/17/2017
• Issued: 05/28/2019
• Est. Priority Date: 03/10/2011
• Status: Active Grant

• Current Assignee: General Atomics, Inc.
• Original Assignee: General Atomics, Inc.
• Inventors: Kelley, Shana O., Bortolin, Susan, Orton, Reginald James McKenzie, Wiechula, Stefan Christopher
• Primary Examiner(s): Crow, Robert T.
• Application Number:
  US15/408,225
• Publication Number:
  US 20170253912A1
• Time in Patent Office: 861 Days
• Field of Search: None
• CPC Class Codes: C12N 1/066 by physical methods
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12N 15/1003 Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2523/307 Denaturation or renaturation by electric current/voltage
• CPC Class Codes: C12Q 2525/204 specific length of the oligonucleotides
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: C12Q 2565/629 being a microfluidic device

Abstract

Contemplated methods and devices are drawn to preparation and analysis of analytes from biological samples. In a preferred embodiment the analytes are nucleic acids that are both released from biological compartments present in the sample and fragmented through the use of a voltage potential applied to a pair of electrodes. The nucleic acids thus prepared are subsequently characterized.

Claims

1. A device for analyzing a biological sample, the device comprising:

a sample receiving zone configured to receive a biological sample, the biological sample including nucleic acids;

a sample extraction zone in fluid communication with the sample receiving zone and configured to receive the biological sample from the sample receiving zone, the sample extraction zone including a plurality of electrodes in electrical communication with an electrical source and configured to fragment the nucleic acids into a plurality of fragmented nucleic acids when a first electrical signal is applied to the plurality of electrodes;

a signal control unit configured to determine an overall treatment time of the first electrical signal applied to the plurality of electrodes based on a desired average length in bases of the plurality of fragmented nucleic acids; and

a sample analysis zone in fluid communication with the sample extraction zone and including a sensing electrode and a reference electrode for analyzing the biological sample.

2. The device of claim 1, further comprising:

a fluid reservoir including a pliant wall defining an interior volume, the pliant wall having an external surface configured such that pressure applied to the external surface of the pliant wall results in deformation of the pliant wall into an interior volume of the fluid reservoir, thereby inducing fluid flow through the sample receiving zone.

3. The device of claim 2, wherein the fluid reservoir includes an extraction buffer.

4. The device of claim 1, further comprising:

a flow channel, wherein each of the plurality of electrodes extends along a length of the flow channel, and wherein each of the plurality of electrodes comprises ridges or projections that protrude into the flow channel.

5. The device of claim 1, wherein each of the plurality of electrodes is disposed in the extraction zone and configured to effect release of a plurality of nucleic acids from the biological sample and to effect fragmentation of the plurality of nucleic acids.

6. The device of claim 1, wherein the sample receiving zone is configured to receive and retain the biological sample and at least a portion of a sample collecting device.

7. The device of claim 1, wherein the sensing electrode includes a probe molecule that is at least partially complementary to the fragmented nucleic acid.

8. The device of claim 7, wherein the sample analysis zone includes a reporter system that is responsive to nucleic acid hybridization to the probe molecule.

9. The device of claim 8, wherein the sensing electrode includes a nanostructured microelectrode, wherein the nanostructured microelectrode is at least one of textured and fractal.