Nucleic acid isolation
View Patent ↗The present invention relates to a method of isolating nucleic acid from a blood sample, said method comprising: (a) selectively isolating leucocytes from said sample by binding said leucocytes to a solid support by means of a binding partner specific for leucocytes; (b) lysing said isolated leucocytes; and (c) binding nucleic acid released from said lysed cells to said solid support. Kits for isolating nucleic acid from samples form further embodiments of the invention.
1. A method of isolating nucleic acid from a blood sample, said method comprising:
(a) selectively isolating leucocytes from said sample by binding said leucocytes to a first solid support by means of at least two different binding partners specific for leucocytes;
(b) lysing said isolated leucocytes;
(c) binding nucleic released from said lysed leucocytes to said first solid support; and
(d) contacting the nucleic acid of step (c) with an additional amount of a second solid support, and binding the nucleic acid to said second solid support;
wherein the method is performed using an automated system for handling the first solid support and second solid support during cell lysis, nucleic acid binding or both, and wherein the first solid support and second solid support are transferred from a first tube to a second tube by the automated system, wherein the at least two different binding partners recognize or are capable of binding specifically to CD45 and CD15, and wherein the first solid support comprises polymeric particles.
2. The method of claim 1 wherein in step (a), said binding partners are attached to a solid support before or after binding to said leucocytes, thereby binding said support to said leucocytes.
3. The method of claim 1 wherein the nucleic acid is DNA, RNA or any naturally occurring modification thereof, or combinations thereof.
4. The method of claim 1 wherein the different binding partners in step (a) bind specifically to leucocytes present in the sample but not to other cells or components of the sample.
5. The method of claim 4 wherein said different binding partners are antibodies or fragments or derivatives of antibodies.
6. The method of claim 1 wherein all or substantially all leucocytes present in the sample are separated.
7. The method of claim 1 wherein said solid support is superparamagnetic.
8. The method of claim 1 wherein the different binding partners are attached directly or indirectly to the solid support of step (a).
9. The method of claim 8 wherein said different binding partners are attached to the same or different solid support.
10. The method of claim 9 wherein the nucleic acid binding in step (c) is carried out using a detergent based system.
11. A method of isolating nucleic acid from a cell sample, said method comprising:
(a) selectively isolating cells from said sample by binding said cells to a first solid support by means of at least two different binding partners specific for said cells;
(b) lysing said isolated cells;
(c) binding nucleic acid released from said lysed cells to said first solid support; and
(d) contacting the nucleic acid of step (c) with an additional amount of a second solid support, and binding said nucleic acid to said second solid support;
wherein the method is performed using an automated system for handling of the first solid support and second solid support during cell lysis, nucleic acid binding or both, and wherein the first solid support and second solid support are transferred from a first tube to a second tube by the automated system, wherein the at least two different binding partners recognize or are capable of binding specifically to CD45 and CD15, and wherein the first solid support comprises polymeric particles.
12. The method of claim 11 wherein the cells are leucocytes.
13. The method of claim 11 wherein the cell sample is a blood sample.
14. The method of claim 11 wherein the first solid support comprises at least two different binding partners that recognize or are capable of binding specifically to one or more of the molecules selected from the group comprising HLA-I, CD11a, CD18, CD45, CD46, CD50, CD82, CD100, CD162, CD5 and CD15 and the second solid support, which is different from said first solid support, is negatively charged.
15. The method of claim 11 wherein said automated system additionally handles the support during the cell isolation stage.
16. The method of claim 11 wherein in step (a), said binding partners attach to a solid support before or after binding to said leucocytes.
17. The method of claim 11 wherein the nucleic acid is DNA, RNA or any naturally occurring modifications thereof, or combinations thereof.
18. The method of claim 11 wherein the different binding partners in step (a) binds specifically to leucocytes present in the sample but not to other cells or components of the sample.
19. The method of claim 11 wherein said different binding partners are antibodies or fragments or derivatives of antibodies.
20. The method of claim 11 wherein all or substantially all leucocytes present in the sample are separated.
21. The method of claim 11 wherein said solid support is superparamagnetic.
22. The method of claim 11 wherein the different binding partners are attached directly or indirectly to the solid support of step (a).
23. The method of claim 22 wherein said different binding partners are attached to the same or different solid support.
24. The method of claim 11 wherein the nucleic acid binding in step (c) is carried out using a detergent based system.
25. The method of claim 11 wherein the nucleic acid of step (c) is additionally contacted with an additional amount of a second solid