Highly simplified lateral flow-based nucleic acid sample preparation and passive fluid flow control

• US 9,207,236 B2
• Filed: 11/05/2010
• Issued: 12/08/2015
• Est. Priority Date: 05/05/2008
• Status: Active Grant

• Current Assignee: TRIAD National Security, LLC
• Original Assignee: Los Alamos National Security LLC (Government of the United States of America)
• Inventors: Cary, Robert B.
• Primary Examiner(s): Chin, Chris L
• Application Number:
  US12/940,973
• Publication Number:
  US 20110117540A1
• Time in Patent Office: 1,859 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C12N 15/101 by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
• CPC Class Codes: G01N 1/405 by adsorption or absorption
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387

Abstract

Highly simplified lateral flow chromatographic nucleic acid sample preparation methods, devices, and integrated systems are provided for the efficient concentration of trace samples and the removal of nucleic acid amplification inhibitors. Methods for capturing and reducing inhibitors of nucleic acid amplification reactions, such as humic acid, using polyvinylpyrrolidone treated elements of the lateral flow device are also provided. Further provided are passive fluid control methods and systems for use in lateral flow assays.

Claims

1. A device for processing a sample comprising a material which defines a flow path and which comprises the following elements:

a sample receiving zone for receiving an aliquot of a fluid sample which comprises targets comprising nucleic acids;

a capture zone in capillary flow contact with said sample receiving zone for binding and/or purifying the targets;

a capillary flow path comprising said material and connecting said sample receiving zone and said capture zone; and

one or more assay zones, each assay zone connected to said capillary flow path via two solution flow paths comprising said material, each said solution flow path forming a junction with said capillary flow path downstream of said sample receiving zone;

each said solution flow path configured to deliver a corresponding solution to said capillary flow path or said capture zone at a desired time or sequence after the aliquot and each solution are all added to the device at the time of assay initiation;

wherein for each assay zone said two junctions are on opposite sides of said capillary flow path; and

wherein the material comprises an absorbent material capable of supporting fluid wicking and the passive control of at least one fluid flow therein.

2. The device according to claim 1, further comprising a lysis zone, in capillary flow contact with said capture zone, which is capable of achieving the lysis of the membrane(s) of a biological particle or cell comprising one or more targets and the liberation of nucleic acid therefrom.

3. The device according to claim 2, wherein the lysis zone comprises a microporous membrane.

4. The device according to claim 3, wherein the microporous membrane comprises nitrocellulose.

5. The device according to claim 2, wherein the lysis zone comprises an absorbent material capable of supporting fluid wicking and the passive control of at least one fluid flow therein.

6. The device according to claim 5 wherein the absorbent material comprises nitrocellulose.

7. The device according to claim 1, wherein said assay zones comprise the nucleic acid and labeling constituents for a sandwich nucleic acid hybridization assay.

8. The device according to claim 1, further comprising a nucleic acid amplification zone downstream of and in capillary flow contact with the capture zone, and upstream and in capillary flow contact with said one or more assay zones.

9. The device according to claim 1, wherein the capture zone comprises a microporous membrane.

10. The device according to claim 9, wherein the microporous membrane comprises nitrocellulose.

11. The device according to claim 1, wherein one or more of the assay zones comprises a microporous membrane.

12. The device according to claim 11, wherein the microporous membrane comprises nitrocellulose.

13. The device according to claim 1, wherein the sample receiving zone and/or capture zone comprises an absorbent material capable of supporting fluid wicking and the passive control of at least one fluid flow therein.

14. The device according to claim 13 wherein the absorbent material comprises nitrocellulose.

15. The device according to claim 1, wherein said capture zone comprises immobilized ligands for binding the targets, silica, glass fiber, and/or diethyl aminoethyl (DEAE) membrane.

16. The device according to claim 1, wherein the absorbent material allows for the passive control of multiple fluid flows therein.

17. The device according to claim 1 wherein the absorbent material comprises nitrocellulose.