Highly Simplified Lateral Flow-Based Nucleic Acid Sample Preparation and Passive Fluid Flow Control

• US 20160083716A1
• Filed: 12/08/2015
• Published: 03/24/2016
• 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.
• Granted Patent: US 9,944,922 B2
• Time in Patent Office: Days
• 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 geometrically-defined nitrocellulose strip having a configuration as shown in FIG. 5, 6, 7, 8, 9 or 14A.

2. A method for measuring an amount of a target, the method comprising the steps of:

disposing a plurality of biological particles comprising one or more targets in a sample receiving zone;

lysing the particles; and

binding the targets to a first ligand in a capture zone in lateral flow connection with the sample receiving zone, thereby increasing a concentration of the target relative to other constituents in the particles.

3. The method of claim 2 wherein the particles are selected from the group consisting of cells, viruses, and bacteria.

4. The method of claim 2 further comprising removing the other constituents from the capture zone.

5. The method of claim 2 further comprising increasing a concentration of the particles prior to the lysing step by reacting a second ligand in the with the surface of the particle.

6. The method of claim 5 wherein the second ligand comprises an antibody or a carbohydrate.

7. The method of claim 2 wherein the binding step comprises binding DNA with silica, binding RNA with silica, or providing a functionalized substrate for anion or cation exchange mediated enrichment of nucleic acids, proteins, or small molecules.

8. The method of claim 22 wherein the targets comprise nucleic acids, and further comprising:

adding a lysate and/or a wash buffer to a buffer exchanger that is in lateral flow contact with a nucleic acid affinity matrix;

and directly purifying the nucleic acids.