Devices, systems and methods for ultra-low volume liquid biopsy
View Patent ↗Provided herein are devices, systems, kits and methods for obtaining genetic information from cell-free fetal nucleic acids in ultra-low amounts of biological samples. Due to the convenience of obtaining ultra-low amounts of samples, devices, systems, kits and methods can be at least partially employed at a point of need.
1. A method comprising:
(a) obtaining or providing capillary blood comprising cell-free nucleic acids obtained from a subject;
(b) extracting the cell-free nucleic acids from the capillary blood;
(c) preparing a nucleic acid library from the cell-free nucleic acids with an efficiency of at least about 0.5, wherein the nucleic acid library obtains the efficiency of at least about 0.5 by using one or more crowding agents;
(d) detecting at least one target nucleic acid sequence present in the nucleic acid library; and
(e) identifying one or more genetic or chromosomal abnormalities based on the detecting of (d).
2. The method of claim 1 , wherein the detecting of (d) comprises sequencing the at least one target nucleic acid sequence.
3. The method of claim 1 , wherein the subject is pregnant with a fetus.
4. The method of claim 3 , wherein the cell-free nucleic acids comprise fetal cell-free nucleic acids.
5. The method of claim 4 , wherein the one or more genetic or chromosomal abnormalities comprises a fetal aneuploidy or a fetal chromosomal abnormality.
6. The method of claim 1 , wherein the subject has, is suspected of having, or is at risk of developing cancer.
7. The method of claim 1 , wherein the capillary blood is obtained from the subject by use of a transdermal puncture device.
8. The method of claim 1 , further comprising separating plasma or serum from the capillary blood or a portion thereof, and the extracting of (b) comprises extracting the cell-free nucleic acids from the plasma or serum.
9. The method of claim 1 , wherein the extracting of (b) comprises binding the cell-free nucleic acids to a solid support.
10. The method of claim 9 , wherein the solid support is selected from the group consisting of: a bead, a nanoparticle, a magnetic particle, a chip, a microchip, a fibrous strip, a polymer strip, a membrane, a matrix, a column, a plate, and any combination thereof.
11. The method of claim 9 , further comprising, eluting the cell-free nucleic acids from the solid support.
12. The method of claim 1 , further comprising, purifying the cell-free nucleic acids.
13. The method of claim 8 , wherein the separating comprises centrifuging the capillary blood, filtering the capillary blood, or both.
14. The method of claim 1 , wherein the detecting of (d) comprises detecting an overrepresentation, an underrepresentation, or a normal representation of the at least one target nucleic acid sequence in the cell-free nucleic acids.
15. The method of claim 1 , wherein a total volume of the capillary blood is from about 5 μL to about 1 mL.
16. The method of claim 1 , further comprising enriching for the cell-free nucleic acids thereby generating enriched cell-free nucleic acids.
17. The method of claim 16 , wherein the enriching comprises removing a first amount of blood from the capillary blood.
18. The method of claim 16 , wherein the enriching comprises enriching for the at least one target sequence present in the cell-free nucleic acids.
19. The method of claim 16 , wherein the enriching comprises enriching for fetal cell-free nucleic acids.
20. The method of claim 16 , wherein the enriching comprises removing white blood cells, cellular nucleic acids, or both, from the capillary blood.
21. The method of claim 1 , wherein the one or more crowding agents is selected from the group consisting of: polyethylene glycol, dextran, and polysaccharide.