Systems and methods for isolating nucleic acids

• US 9,803,188 B2
• Filed: 12/21/2012
• Issued: 10/31/2017
• Est. Priority Date: 12/22/2011
• Status: Expired due to Fees

• Current Assignee: Ibis Biosciences Incorporated (Abbott Laboratories)
• Original Assignee: Ibis Biosciences Incorporated (Abbott Laboratories)
• Inventors: Eshoo, Mark W., Crowder, Christopher
• Primary Examiner(s): THOMAS, DAVID C
• Application Number:
  US14/367,796
• Publication Number:
  US 20140323319A1
• Time in Patent Office: 1,775 Days
• Field of Search: None
• CPC Class Codes: C12N 15/1013 by using magnetic beads
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/701 Specific hybridization probes

Abstract

The present disclosure relates to systems and methods for nucleic acid isolation. In particular, the present disclosure provides systems and methods for isolating nucleic acids from aqueous samples (e.g., blood or urine).

Claims

1. A method of simultaneously extracting DNA and RNA from a sample, comprising:

a) disrupting cells in said sample;

then

b) contacting said sample comprising said disrupted cells with a lysis solution comprising isopropanol to generate a mixture of said sample comprising said disrupted cells, said lysis solution and said isopropanol;

then

c) mixing said mixture;

then

d) incubating said mixture to generate a lysed sample; and

then

e) extracting nucleic acids from said lysed sample to generate extracted nucleic acids.

2. The method of claim 1, wherein said sample is an aqueous biological fluid.

3. The method of claim 2, wherein said aqueous biological fluid is selected from the group consisting of blood, blood products, and urine.

4. The method of claim 1, wherein said incubating comprises incubating at 30°

C. for 1 hour.

5. The method of claim 1, further comprising the step of analyzing said extracted nucleic acids from said sample for the presence of a target DNA and/or RNA.

6. The method of claim 5, wherein said target nucleic acid is a pathogen nucleic acid.

7. The method of claim 6, wherein said pathogen is selected from the group consisting of a virus, a fungi, and a bacterium.

8. The method of claim 5, wherein said analyzing comprises a method selected from the group consisting of an amplification method, a sequencing method, and a hybridization method.

9. The method of claim 1, wherein said disrupting comprises bead beating in the presence of a detergent.

10. The method of claim 1, wherein said extracting step e) comprises binding said nucleic acids to a bead or solid support.

11. The method of claim 1, wherein said method is automated.

12. The method of claim 1, wherein said disrupting comprises a proteinase.

13. The method of claim 1, wherein said isopropanol is 15% isopropanol.

14. A method of simultaneously extracting DNA and RNA from a sample, comprising:

a) bead beating said sample in the presence of a detergent and/or a proteinase to generate disrupted cells in said sample;

then

b) contacting said sample comprising said disrupted cells with isopropanol;

then

c) contacting said sample comprising said disrupted cells with said isopropanol with a lysis solution;

then

d) mixing said sample comprising said disrupted cells with said isopropanol with said lysis solution;

then

e) incubating said sample comprising said disrupted cells with said isopropanol with said lysis solution at 30°

C. for one hour to generate a lysed sample;

then

f) extracting DNA and RNA from said lysed sample to generate extracted DNA and RNA.