Apparatus for point-of-care detection of nucleic acid in a sample

Patent metadata

Field	Value
Publication number	US-9315858-B2
Application number	US-201113115877-A
Country	US
Kind code	B2
Filing date	May 25, 2011
Priority date	May 25, 2010
Publication date	Apr 19, 2016
Grant date	Apr 19, 2016

Abstract

Provided herein are methods and apparatus for detecting a target nucleic acid in a sample and related methods and apparatus for diagnosing a condition in an individual. The condition is associated with presence of nucleic acid produced by certain pathogens in the individual.

First claim

What is claimed is:

1. Apparatus for detecting a target nucleic acid in a sample, the apparatus comprising:
a containment vessel being a sealable tube, wherein the containment vessel has a container body defining a single unpartitioned cavity;
a sample collection element configured for removable coupling to the containment vessel;
a nucleic acid binding element positioned within the containment vessel,
wherein the sample collection element is configured to collect the sample and to transfer the sample to the nucleic acid binding element when the sample collection element is removably coupled to the containment vessel, and
wherein the sample collection element includes a lumen and a swab at one end of the lumen, the lumen being configured to deliver through the swab a fluid to the containment vessel;

a plurality of reagents configured for placement in fluid communication with the nucleic acid binding element,
wherein the plurality of reagents comprises nucleic acid purification reagents and nucleic acid amplification reagents; and

a heater configured to heat the nucleic acid amplification reagents in fluid communication with the nucleic acid binding element, the heater being a disposable heater applied on an external surface around the containment vessel.

2. The apparatus of claim 1 , wherein the containment vessel is sealed when the sample collection element is removably coupled to the containment vessel.

3. The apparatus of claim 1 ,
wherein the containment vessel comprises a loading port, and
wherein the plurality of reagents are configured for loading or extracting through the loading port for placement in or removal from fluid communication with the nucleic acid binding element.

4. The apparatus of claim 1 ,
wherein the sample collection element comprises a lumen, and
wherein the sample collection element is configured to transfer the sample to the nucleic acid binding element via lavaging the nucleic acid purification reagents through the lumen when the sample collection element is removably coupled to the containment vessel.

5. The apparatus of claim 4 , wherein the plurality of reagents are configured for loading through the lumen for placement in fluid communication with the nucleic acid binding element.

6. The apparatus of claim 1 , wherein the nucleic acid amplification reagents are suitable for isothermal amplification of the target nucleic acid.

7. The apparatus of claim 6 , wherein the isothermal amplification is selected from the group consisting of loop-mediated isothermal amplification, helicase-dependent isothermal amplification and recombinase polymerase amplification.

8. The apparatus of claim 1 , wherein the containment vessel, the sample collection element, the nucleic acid binding element, and the plurality of reagents are configured for a single use.

9. The apparatus of claim 1 , wherein the heater is configured for a single use.

10. The apparatus of claim 1 , further comprising a waste collection unit.

11. The apparatus of claim 10 , wherein the waste collection unit is coupled to the sample collection element and comprises:
a chamber, the chamber comprising a first gap; and
a plate, the plate coupled with the chamber and comprising a second gap,
wherein the chamber and the plate surround the sample collection element and at least one of the chamber and the plate is configured to be rotatable, and
wherein when the first gap overlaps with the second gap, content of the containment vessel is adapted to flow into the chamber of the waste collection unit.

12. The apparatus of claim 11 , wherein the content of the containment vessel adapted to flow into the chamber of the waste collection unit is a waste.

13. The apparatus of claim 5 , wherein the plurality of reagents are enveloped within a plurality of reagent cartridges,
wherein the plurality of reagent cartridges each comprises a capsule and a plug, the capsule being connected to the lumen through an opening, the plug sealing the opening; and
wherein the plurality of reagent cartridges are configured to load the plurality of reagents through the lumen for placement in fluid communication with the nucleic acid binding element upon removal of the plug from the opening.

14. The apparatus of claim 1 , wherein the nucleic acid amplification reagents are sequestered within separate polymer shells, the polymer shells comprised within the containment vessel and configured to melt and to release the nucleic acid amplification reagents for placement in fluid communication with the nucleic acid binding element when the polymer shells are heated by the heater.

15. The apparatus of claim 14 , wherein the polymer shells are polycaprolactone (PCL) shells.

16. The apparatus of claim 1 , wherein the sample collection element comprises a bodily sample collection element suitable for collecting a bodily sample selected from the group consisting of nasal mucus, urine, fecal matter, blood, saliva, buccal cells and combinations thereof.

17. The apparatus of claim 16 , wherein the bodily sample collection element is a swab.

18. The apparatus of claim 1 , wherein the nucleic acid binding element comprises a FTA card.

19. The apparatus of claim 1 , wherein the heater is configured to generate heat via an exothermal chemical reaction.

20. The apparatus of claim 19 , wherein the heater comprises a phase change material.

21. The apparatus of claim 1 , wherein the heater is configured to generate heat via metal oxidation.

22. The apparatus of claim 1 , wherein the heater is configured to generate heat using a supersaturated salt solution.

23. The apparatus of claim 1 , further comprising a detection element configured to detect amplification of the target nucleic acid.

24. The apparatus of claim 23 ,
wherein the detection element comprises a colorimetric dye in fluid communication with the nucleic acid amplification reagents, and
wherein the colorimetric dye is configured to undergo a color shift when the target nucleic acid is amplified.

25. The apparatus of claim 24 , wherein the colorimetric dye is hydroxynaphthol blue (HNB).

26. The apparatus of claim 23 ,
wherein the detection element comprises a fluorescent dye in fluid communication with the nucleic acid amplification reagents, and
wherein the fluorescent dye is configured to fluoresce when the target nucleic acid is amplified.

27. Apparatus for diagnosing a condition in an individual, the condition being associated with presence of a target nucleic acid in the individual, the target nucleic acid being produced by a pathogen, the apparatus comprising:
a containment vessel being a sealable tube, wherein the containment vessel has a container body defining a single unpartitioned cavity;
a sample collection element configured for removable coupling to the containment vessel and comprising a lumen;
a nucleic acid binding element positioned within the containment vessel,
wherein the sample collection element is configured to collect the sample and to transfer the sample to the nucleic acid binding element when the sample collection element is removably coupled to the containment vessel, and
wherein the sample collection element includes a lumen and a swab at one end of the lumen, the lumen being configured to deliver through the swab a fluid to the containment vessel;

a waste collection unit comprising a chamber and a plate, the chamber co

Assignees

• Bearinger Jane P
• Dugan Lawrence C
• L Livermore Nat Security Llc

Inventors

• Bearinger Jane P
• Dugan Lawrence C

Classifications

• C12Q1/6844 Primary

  Nucleic acid amplification reactions · CPC title

• C12Q1/689

  for bacteria · CPC title

• C12Q1/701

  Specific hybridization probes · CPC title

• B01L2300/0851

  Bottom walls · CPC title

• B01L3/502776

  specially adapted for focusing or laminating flows · CPC title