Methods For Detection Of Sepsis

• US 20100160171A1
• Filed: 12/22/2009
• Published: 06/24/2010
• Est. Priority Date: 12/22/2008
• Status: Abandoned Application

• Current Assignee: Children's National Medical Center
• Original Assignee: Children's Research Institute
• Inventors: FREISHTAT, Robert J.
• Application Number:
  US12/644,901
• Publication Number:
  US 20100160171A1
• Time in Patent Office: Days
• US Class Current: 506/7
• CPC Class Codes: C12Q 1/37 involving peptidase or proteinase
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/142 Toxicological screening, e.g. expression profiles which identify toxicity
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 2333/96436 Granzymes
• CPC Class Codes: G01N 2800/26 Infectious diseases, e.g. generalised sepsis
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Abstract

The present invention relates to a method for diagnosis, detection, or prognosis of sepsis and its severity. More specifically, this invention uses the presence and amount of granzyme B in platelets as a marker for sepsis.

Claims

1. A method for detecting or diagnosing or monitoring the progression of sepsis in an individual comprising the step of determining presence of granzyme B in platelets of the individual.

2. The method of claim 1, wherein the determining step is accomplished by immunoassay.

3. The method of claim 2, wherein the immunoassay is ELISA.

4. The method of claim 2, wherein in the immunoassay is an immunoblot.

5. The method of claim 1, wherein the determining step is accomplished by measuring nucleic acid levels.

6. The method of claim 4, wherein the nucleic acid is mRNA.

7. The method of claim 6, wherein the mRNA codes for granzyme B.

8. The method of claim 6, wherein the nucleic acid levels are measured by Northern blot.

9. The method of claim 6, wherein the nucleic acid levels are measured by microarray analysis.

10. The method of claim 1, wherein the determining step comprises the steps of

contacting a sample from the individual with a molecule that specifically binds the granzyme B;

and

detecting the presence of binding between the granzyme B and the molecule.

11. The method of claim 10, wherein the molecule is an antibody.

12. The method of claim 11, wherein the antibody is selected from the group consisting of monoclonal antibodies and polyclonal antibodies.

13. The method of claim 10, wherein the molecule is labeled.

14. The method of claim 13, wherein the label is selected from the group consisting of biotin, fluorescent molecules, radioactive molecules, chromogenic substrates, chemiluminescence, and enzymes.

15. The method of claim 1, wherein the determining step comprises the steps of

isolating mRNA from the platelets of the individual;

contacting the isolated mRNA with a probe that specifically hybridizes with the mRNA of the granzyme B; and

detecting the presence of binding between the probe and the mRNA.

16. The method of claim 15, wherein the probe is a nucleic acid probe.

17. The method of claim 16, wherein the probe is an oligonucleotide.

18. The method of claim 16, wherein the probe is labeled.

19. The method of claim 18, wherein the label is selected from the group consisting of biotin, fluorescent molecules, radioactive molecules, chromogenic substrates, chemiluminescence, and enzymes.

20. The method of claim 15, wherein the probe is attached to a solid substrate.

21. The method of claim 15, wherein the probe is on a microarray.

22. A method for monitoring the treatment of an individual with sepsis comprising the steps of

administering a composition for treating sepsis to the individual;

and

determining presence of granzyme B in platelets of the individual.

23. The method of claim 22, wherein the determining step is accomplished by immunoassay.

24. The method of claim 23, wherein the immunoassay is ELISA.

25. The method of claim 24, wherein in the immunoassay is an immunoblot.

26. The method of claim 22, wherein the determining step is accomplished by measuring nucleic acid levels.

27. The method of claim 26, wherein the nucleic acid is mRNA.

28. The method of claim 27, wherein the mRNA codes for granzyme B.

29. The method of claim 25, wherein the nucleic acid levels are measured by Northern blot.

30. The method of claim 25, wherein the nucleic acid levels are measured by microarray analysis.

31. The method of claim 24, wherein the determining step comprises the steps of

contacting the serum of the individual with a molecule that specifically binds the granzyme B;

and

detecting a presence of binding between the granzyme B and the molecule.

32. The method of claim 31, wherein the molecule is an antibody.

33. The method of claim 32, wherein the antibody is selected from the group consisting of monoclonal antibodies and polyclonal antibodies.

34. The method of claim 31, wherein the molecule is labeled.

35. The method of claim 34, wherein the label is selected from the group consisting of biotin, fluorescent molecules, radioactive molecules, chromogenic substrates, chemiluminescence, and enzymes.

36. The method of claim 22, wherein the determining step comprises the steps of

isolating mRNA from the platelets;

contacting the isolated mRNA with a probe that specifically hybridize with the mRNA of the granzyme B; and

detecting a presence of binding between the probe and the mRNA.

37. The method of claim 36, wherein the probe is a nucleic acid probe.

38. The method of claim 37, wherein the probe is an oligonucleotide.

39. The method of claim 37, wherein the probe is labeled.

40. The method of claim 39, wherein the label is selected from the group consisting of biotin, fluorescent molecules, radioactive molecules, chromogenic substrates, chemiluminescence, and enzymes.

41. The method of claim 38, wherein the probe is attached to a solid substrate.

42. The method of claim 38, wherein the probe is on a microarray.

43. The method of claim 22, further comprising the step of comparing the presence of granzyme B of the individual over time to determine the effect to the composition on the progression of the sepsis.