Protein-based sample collection matrices and devices

Patent metadata

Field	Value
Publication number	US-2017327815-A1
Application number	US-201715592481-A
Country	US
Kind code	A1
Filing date	May 11, 2017
Priority date	May 13, 2016
Publication date	Nov 16, 2017
Grant date	—

Abstract

Provided herein are methods and compositions to effectively isolate nucleic acids using protein-based sample collection matrices.

First claim

We claim:

1 . A kit for isolating nucleic acids from a sample, the kit comprising:
a) a protein-based matrix configured to collect sample; b) a solution comprising about 3-6 M chaotrope, about 0.5-10% detergent, about 50-500 mM reducing agent, and buffer maintaining pH of the solution at about 5-8.

2 . The kit of claim 1 , wherein the protein-based matrix forms a swab.

3 . The kit of claim 1 , wherein the protein-based matrix forms a strip to collect liquid sample.

4 . The kit of claim 1 , further comprising processing vessels.

5 . The kit of claim 1 , further comprising filter tubes or magnetic glass beads (MGPs).

6 . The kit of claim 5 , further comprising wash buffer and elution buffer.

7 . The kit of claim 5 , wherein the protein-based matrix is silk.

8 . The kit of claim 5 , wherein the chaotrope is a guanidinium salt.

9 . The kit of claim 5 , wherein the detergent is a non-ionic detergent.

10 . The kit of claim 5 , wherein the reducing agent is dithiothreitol.

11 . The kit of claim 5 , wherein the buffer is a citrate buffer.

12 . The kit of claim 5 , wherein the pH of the solution is 6-7.

13 . The kit of claim 5 , further comprising a container for storing the protein-based matrix and sample.

14 . A method for collecting nucleic acids in a sample, the method comprising
contacting a sample containing nucleic acids on a protein-based matrix, thereby collecting nucleic acids.

15 . The method of claim 14 , further comprising placing the protein-based matrix in a container for storage.

16 . The method of claim 14 , further comprising contacting the protein-based matrix with a solution comprising about 3-6 M chaotrope, about 0.5-10% detergent, about 50-500 mM reducing agent, and buffer maintaining pH of the solution at about 5-8, thereby solubilizing the protein-based matrix in the solution.

17 . The method of claim 16 , wherein the protein-based matrix is silk.

18 . The method of claim 16 , wherein the chaotrope is a guanidinium salt.

19 . The method of claim 16 , wherein the detergent is a non-ionic detergent.

20 . The method of claim 16 , wherein the reducing agent is dithiothreitol.

21 . The method of claim 16 , wherein the buffer is a citrate buffer.

22 . The kit of claim 16 , wherein the pH of the solution is 6-7.

23 . The method of claim 16 , wherein the protein-based matrix is immersed in the solution for at least 10 minutes.

24 . The method of claim 16 , further comprising contacting the solution with a filter tube or with magnetic glass beads (MGPs), washing the filter tube or MGPs with wash buffer, and eluting the nucleic acids from the filter tube or MGPs with elution buffer, thereby isolating the nucleic acids from the sample.

25 . The method of claim 16 , wherein the sample is a liquid sample.

Assignees

• Roche Molecular Systems Inc

Inventors

• Will Stephen

Classifications

• G01N2001/028

  Sampling from a surface, swabbing, vaporising · CPC title

• C12Q1/6806

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• G01N1/10

  in the liquid or fluent state {(burettes, pipettes B01L3/02; sampling of ground water E02D1/06; metering by volume of fluids or fluent solid material G01F11/00, G01F13/00)} · CPC title

• C12N15/1006 Primary

  by means of a solid support carrier, e.g. particles, polymers · CPC title

• A61B10/02

  Instruments for taking cell samples or for biopsy {(A61B10/0038 and A61B10/0045 take precedence; needle locating or guiding means A61B17/3403; samplers for enzymology or microbiology C12M1/26; sampling or preparing biological specimens G01N33/48)} · CPC title