Isolation of cell-free nucleic acids from bodily fluid samples using solid chaotropic agents

1. A process for isolating cell-free nucleic acid or an analog thereof from a bodily fluid, comprising: a) mixing in a container the bodily fluid, a chaotropic agent in solid form, a detergent in solid form and a buffer in solid form, and a solid phase which comprises magnetic particles, thus forming a reaction mixture containing the cell-free nucleic acid, wherein the reaction mixture has a volume of about 2 ml to about 10 ml, and wherein the chaotropic agent is present in the reaction mixture in an amount of about 2 to about 10 grams and at a concentration of about 3M to about 10M; b) magnetically separating the solid phase having the cell-free nucleic acid bound thereto from the reaction mixture; and optionally c) dissociating the nucleic acid from the solid phase.

2. A process according to claim 1 , wherein the bodily fluid comprises blood or a component thereof.

3. A process according to claim 2 , wherein the blood component is plasma.

4. A process according to claim 2 , wherein the blood component is serum.

5. A process according to claim 1 , wherein the bodily fluid comprises urine.

6. A process according to claim 1 , wherein the bodily fluid sample is obtained from a pregnant female.

7. A process according to claim 6 , wherein the bodily fluid is maternal blood or a component thereof.

8. A process according to claim 1 , wherein the bodily fluid is obtained from a cancer patient who has been treated with a therapeutic agent which comprises a nucleic-acid analog.

9. A process according to claim 8 , wherein the nucleic acid analog is a peptide nucleic acid (PNA), a phosphorothioate, locked nucleic acid (LNA), or an oligonucleotide or a chemically modified oligonucleotide.

10. A process according to claim 1 , wherein the chaotropic agent is selected from the group consisting of a guanidinium salt, an iodide, sodium thiocyanate and urea.

11. A process according to claim 10 , wherein the guanidinium salt is guanidinium (iso)thiocyanate.

12. A process according to claim 1 , wherein the nucleic acid is DNA.

13. A process according to claim 1 , wherein the nucleic acid is RNA.

14. A process according to claim 1 , wherein the solid phase further comprises silica particles having a particle size ranging substantially between 0.05 and 0.4 μm.

15. A process according to claim 1 , further comprising washing the solid phase having the cell-free nucleic acid bound thereto with a washing buffer optionally containing a chaotropic agent.

16. A process according to claim 15 , further comprising washing the thus-washed solid phase with an aqueous solution comprising a monohydric alcohol or a polyol.

17. A process according to claim 16 , wherein the nucleic acid is dissociated from the solid phase by an aqueous elution buffer.

18. A process according to claim 1 , wherein the reaction mixture has a volume of about 2 ml to about 10 ml.

19. A composition useful for isolating cell-free nucleic acid from a bodily fluid, comprising a chaotropic agent in solid form, a buffer in solid form, and a detergent in solid form, wherein the chaotropic agent is present in an amount of about 2 to about 10 grams and is selected such that when mixed with a volume of sample fluid to form a reaction mixture having a volume of about 2 to about 10 milliliters, the chaotropic agent is present in a concentration of about 3M to about 10M.

20. A composition according to claim 19 , wherein the buffer is TRIS HCl.

21. A composition according to claim 19 , wherein the detergent is sodium deoxycholate.

22. A composition according to claim 19 , wherein the detergent is sodium dodecyl sulfate (SDS).

23. A composition according to claim 19 , further comprising magnetic particles.

24. A composition according to claim 19 , further comprising a chelating agent or an anti-clotting agent, or a combination thereof.

25. A test kit for isolating cell-free nucleic acid from a bodily fluid, comprising in a first container, a chaotropic agent in solid form in an amount of about 2 to about 10 grams and is selected such that when mixed with a volume of sample fluid to form a reaction mixture having a volume of about 2 to about 10 milliliters, the chaotropic agent is present in a concentration of about 3M to about 10M, a buffer in solid form, and a detergent in solid form.

26. A test kit according to claim 25 , wherein the chaotropic agent is guanidine thiocyanate, and the detergent is SDS.

27. A test kit according to claim 25 , wherein the first container further comprises a solid phase comprising magnetic particles.

28. A test kit according to claim 25 , further comprising a second container having disposed therein a solid phase comprising magnetic particles.

29. A test kit according to claim 27 , wherein the magnetic particles are coated with silica.

30. A test kit according to claim 25 , further comprising a second container having disposed therein a washing buffer comprising a chaotrope.

31. A test kit according to claim 25 , further comprising a third container comprising a washing buffer comprising a drying agent.

32. A test kit according to claim 25 , further comprising a fourth container comprising an elution buffer.

33. A test kit according to claim 25 , comprising a first container having disposed therein the chaotrope in solid form in an amount of about 2 to about 10 grams and is selected such that when mixed with a volume of sample fluid to form a reaction mixture having a volume of about 2 to about 10 milliliters, the chaotropic agent is present in a concentration of about 3M to about 10M, the solid buffer and the solid detergent; a second container having disposed therein a solid phase comprising magnetic particles coated with silica; and a third container comprising a washing buffer comprising ethanol.

34. A test kit according to claim 28 , wherein the magnetic particles are coated with silica.

35. A composition according to claim 19 , wherein the chaotropic agent is present in an amount of about 4 to about 6 grams.

36. A test kit according to claim 25 , wherein the chaotropic agent is present in an amount of about 4 to about 6 grams.

37. A composition according to claim 19 , wherein the chaotropic agent is present in an amount effective to achieve a molarity of about 4M to about 6M when mixed with a volume of sample fluid of about 2 to about 10 milliliters.

38. A composition according to claim 19 , wherein the chaotropic agent is present in an amount effective to achieve a molarity of about 4M to about 7.5M when mixed with a volume of sample fluid of about 2 to about 10 milliliters.

39. A composition according to claim 19 , wherein the chaotropic agent is present in an amount effective to achieve a molarity of about 5.5M to about 6.5M when mixed with a volume of sample fluid of about 2 to about 10 milliliters.

40. A composition according to claim 19 , wherein the chaotropic agent is present in an amount effective to achieve a molarity of about 3M to about 10M when mixed with a volume of sample fluid of about 2.5 to about 5 milliliters.

41. A composition according to claim 19 , wherein the chaotropic agent is present in an amount effective to achieve a molarity of about 3M to about 10M when mixed with a volume of sample fluid of about 3 to about 5 milliliters.

42. A composition according to claim 19 , wherein the chaotropic agent is present in an amount effective to achieve a molarity of about 3M to about 10M when mixed with a volume of sample fluid of about 5 milliliters.

43. A composition according to claim 19 , wherein the chaotropic agent is present in an amount effective to achieve a molarity of about 5.5M to about 6.5M when mixed with a volume of sample fluid of about 3 to about 5 milliliters.