Compositions and methods for nucleic acid extraction

Patent metadata

Field	Value
Publication number	US-10072259-B2
Application number	US-201615212815-A
Country	US
Kind code	B2
Filing date	Jul 18, 2016
Priority date	Mar 15, 2013
Publication date	Sep 11, 2018
Grant date	Sep 11, 2018

Abstract

Compositions and techniques for the extraction, enrichment and isolation of nucleic acids from yeast in a whole blood sample using amine monomers are disclosed herein.

First claim

I claim:

1. A method of isolating an extraction solution comprising nucleic acids from yeast in a whole blood sample, said method comprising:
a) providing i) a whole blood sample suspected of comprising yeast and ii) an aqueous extraction solution comprising an amine monomer, a chaotrope, and a detergent, wherein said amine monomer is selected from the group consisting of is 2,2′-(ethylenedioxy)bis(ethylamine) (EDBE), 1,3-diaminopropane and 3-amino-1-propanol and wherein the concentration of amine monomer in said aqueous extraction solution is about 30% to about 50% and wherein the concentration of said detergent is about 8% to about 15%;
b) contacting said whole blood sample suspected of comprising yeast with said extraction solution resulting in lysis of the yeast and the isolation of an extraction solution comprising nucleic acids.

2. The method of claim 1 , wherein said amine monomer is a primary amine monomer.

3. The method of claim 2 , wherein said amine monomer is 2,2′-(ethylenedioxy)bis(ethylamine) (EDBE).

4. The method of claim 2 , wherein said amine monomer is 1,3-diaminopropane.

5. The method of claim 2 , wherein said amine monomer is 3-amino-1-propanol.

6. The method of claim 1 , wherein said chaotrope is selected from at least one of the group consisting of urea, and guanidine thiocyanate (GITC).

7. The method of claim 6 , wherein the concentration of the chaotrope in said aqueous extraction solution is about 4 M to about 5 M.

8. The method of claim 1 , wherein said detergent is selected from at least one of the group consisting of a polysorbate, deoxycholate, sodium deoxycholate, sodium dodecyl sulfate (SDS), nonyl phenoxypolyethoxylethanol and polyethylene glycol.

9. The method of claim 1 , further comprising an alcohol wherein said alcohol is selected from at least one of the group consisting of ethanol and butanol.

10. The method of claim 9 , wherein the concentration of said alcohol is about 15% to about 25%.

11. The method of claim 1 , wherein said sample is selected from living cellular source material and fixed cellular source material.

12. The method of claim 11 , wherein said living cellular source material comprises a suspension of single cells.

13. The method of claim 11 , wherein said sample comprises formalin-fixed paraffin embedded (FFPE) material.

14. The method of claim 1 , further comprising wherein said aqueous extraction solution is enzyme-free.

15. The method of claim 1 , further comprising wherein said aqueous extraction solution is protease-free.

16. The method of claim 1 , further comprising wherein said nucleic acids are isolated from an extraction solution comprising nucleic acids with additional steps, the additional steps comprising i) adsorbing said nucleic acids onto silica coated magnetic particles by introducing said particles into said extraction solution containing extracted nucleic acids to create nucleic acid adsorbed silica coated magnetic particles; ii) washing said nucleic acid adsorbed silica coated magnetic particles, iii) eluting said nucleic acids from said silica coated magnetic particles into the elution solution and; iv) isolating said nucleic acids from said elution solution.

17. The method of claim 1 , wherein said method excludes mechanical disruption of the yeast.

Assignees

• Abbott Molecular Inc

Inventors

• Gundling Gerard J

Classifications

• C07H1/08

  from natural products · CPC title

• C07H19/16

  Purine radicals · CPC title

• C12Q1/6806

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

• C12N15/1003 Primary

  Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor · CPC title

• C12N15/1013

  by using magnetic beads · CPC title