SYSTEMS AND METHODS FOR ISOLATING NUCLEIC ACIDS

• US 20140272968A1
• Filed: 03/13/2014
• Published: 09/18/2014
• Est. Priority Date: 03/13/2013
• Status: Active Grant

• Current Assignee: Abbott Molecular Incorporated (Abbott Laboratories Incorporated)
• Original Assignee: Abbott Molecular Incorporated (Abbott Laboratories Incorporated)
• Inventors: Gundling, Gerard, Kowal, Robert, Granados, Edward, Solomon, Natalie, Mak, Wai-Bing, Szostak, Magdalena
• Granted Patent: US 10,640,808 B2
• Time in Patent Office: Days
• US Class Current: 435/6.11
• CPC Class Codes: C12N 1/06 Lysis of microorganisms
• CPC Class Codes: C12N 15/1003 Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence

Abstract

The present disclosure relates to systems and methods for nucleic acid isolation. In particular, the present disclosure provides systems and methods for isolating low molecular weight circulating nucleic acids from bodily fluids (e.g., plasma).

Claims

1. A method of isolating nucleic acids, comprising:

a) contacting a sample comprising nucleic acids with a buffer comprising greater than 35% ethanol by volume; and

b) isolating said nucleic acid from said sample.

2. The method of claim 1, wherein said ethanol is present in said buffer at a concentration of approximately 40% to 60%.

3. The method of claim 1, wherein said ethanol is present in said buffer at a concentration of approximately 45% to 55%.

4. The method of claim 1, wherein said ethanol is present in said buffer at a concentration of approximately 50%.

5. The method of claim 1, wherein said isolating said nucleic acid comprises the steps of i) binding said nucleic acid to a solid support;

ii) washing said solid support with a wash buffer; and

iii) eluting said nucleic acids from said solid support.

6. The method of claim 1, wherein said nucleic acid is a circulating DNA.

7. The method of claim 6, wherein said DNA is less than 1000 bases in length.

8. The method of claim 6, wherein said DNA is less than 500 bases in length.

9. The method of claim 6, wherein said DNA is less than 200 bases in length.

10. The method of claim 1, wherein said sample is selected from the group consisting of blood, blood products, serum, and urine.

11. The method of claim 1, wherein said sample is from a subject and the presence of said nucleic acid in said sample is indicative of a disease state in said subject.

12. The method of claim 11, wherein said disease state is cancer.

13. The method of claim 1, further comprising the step of analyzing said sample for the presence of said nucleic acid.

14. The method of claim 13, wherein said analyzing comprises performing a nucleic acid detection assay selected from the group consisting of an amplification assay, a hybridization assay, a methylation status detection assay, and a sequencing assay.

15. The method of claim 14, wherein said amplification is real time PCR and said methylation status detection assay is methylation specific PCR or heavy methyl PCR.

16. A kit, comprising:

a) a buffer comprising 35% or more ethanol by volume;

b) a wash buffer

c) a solid support; and

d) an elution buffer.

17. The kit of claim 16, wherein said ethanol is present in said buffer at a concentration of approximately 40% to 60%.

18. The kit of claim 16, wherein said ethanol is present in said buffer at a concentration of approximately 45% to 55%.

19. A composition, comprising:

a circulating DNA; and

a buffer comprising 35% or more ethanol by volume.

20. The composition of claim 19, wherein said ethanol is present in said buffer at a concentration of approximately 45% to 55%.