Nucleic acid preparation compositions and methods

• US 10,053,685 B2
• Filed: 11/15/2017
• Issued: 08/21/2018
• Est. Priority Date: 04/03/2009
• Status: Active Grant

• Current Assignee: Sequenom Incorporated (Laboratory Corporation Of America Holdings)
• Original Assignee: Sequenom Incorporated (Laboratory Corporation Of America Holdings)
• Inventors: Wisniewski, Michele Elizabeth, Kwong, William Hang, Mohsenian, Firouz, Ding, Jian-Hua
• Primary Examiner(s): Martinell, James
• Application Number:
  US15/813,979
• Publication Number:
  US 20180073010A1
• Time in Patent Office: 279 Days
• Field of Search: None
• 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: C12N 15/1006 by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2565/137 Chromatographic separation
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Provided herein are methods and compositions to extract and enrich by, physical separation or amplification, relatively short nucleic acids from a nucleic acid composition containing a high background of longer nucleic acids (e.g., host or maternal nucleic acids; genomic nucleic acid and the like).

Claims

1. A method for enriching relatively short nucleic acid from a nucleic acid composition, which comprises:

(a) contacting cell-free nucleic acid from a substantially cell-free biological composition with a solid phase under association conditions, wherein;

(i) the cell-free nucleic acid of the substantially cell-free biological composition comprises relatively short nucleic acid and relatively long nucleic acid,

(ii) the relatively short nucleic acid is about 300 base pairs or less;

(iii) the relatively long nucleic acid is larger than about 300 base pairs, and

(iv) the association conditions do not include polyethylene glycol;

(b) introducing the solid phase after (a) to dissociation conditions that comprise a salt, wherein;

(i) the salt is not a chaotropic salt, and

(ii) the relatively short nucleic acid dissociates from the solid phase under the dissociation conditions, thereby yielding dissociated nucleic acid; and

(c) separating the dissociated nucleic acid from the solid phase, whereby the relatively short nucleic acid is enriched in the dissociated nucleic acid relative to the cell-free nucleic acid from a substantially cell-free biological composition.

2. The method of claim 1, wherein the substantially cell-free biological composition is from a pregnant female.

3. The method of claim 2, wherein the substantially cell-free biological composition is blood serum.

4. The method of claim 2, wherein the substantially cell-free biological composition is blood plasma.

5. The method of claim 2, wherein the substantially cell-free biological composition is urine.

6. The method of claim 2, which further comprises detecting the presence or absence of fetal nucleic acid.

7. The method of claim 6, which further comprises detecting the presence or absence of a chromosome abnormality.

8. The method of claim 7, wherein the chromosome abnormality is a trisomy and the trisomy is trisomy 21, trisomy 18, trisomy 13 or combination thereof.

9. The method of claim 1, wherein the solid phase is a collection of particles.

10. The method of claim 9, wherein the particles comprise silica.

11. The method of claim 1, wherein the solid phase does not comprise a functional group that interacts with the nucleic acid.

12. The method of claim 1, wherein the solid phase has a net charge.

13. The method of claim 1, wherein the dissociation conditions comprise a volume exclusion agent.

14. The method of claim 13, wherein the volume exclusion agent comprises a polyalkyl alcohol, dextran, Ficoll, polyvinyl pyrollidone or combinations thereof.

15. The method of claim 1, wherein the salt comprises an ionic salt.

16. The method of claim 15, wherein the ionic salt is sodium chloride.

17. The method of claim 1, wherein the dissociated nucleic acid comprises deoxyribonucleic acid (DNA).

18. The method of claim 1, wherein the relatively short nucleic acid is about 200 base pairs or less and the relatively long nucleic acid is larger than about 200 base pairs.

19. The method of claim 18, wherein the relatively short nucleic acid is about 50 to about 180 base pairs.

20. The method of claim 1, which further comprises washing the solid phase after (a).

21. The method of claim 1, wherein the solid phase is paramagnetic and the dissociated nucleic acid is separated from the solid phase by a magnet.

22. The method of claim 1, wherein the relatively short nucleic acid is enriched about 10% to about 45% in the dissociated nucleic acid relative to the cell-free nucleic acid from a substantially cell-free biological composition.

23. The method of claim 1, wherein the solid phase is separated from the dissociated nucleic acid by transferring the dissociated nucleic acid to an environment that does not contain the solid phase used in (a).

24. The method of claim 23, wherein the environment is a vessel.