METHODS OF DIAGNOSING BRAIN INJURY

Abstract

Disclosed herein are methods and systems of determining whether a subject’s levels of GFAP, UCH-L1, or GFAP and UCH-L1 are elevated in a sample collected from the subject. The methods comprise determining whether the levels of GFAP, UCH-L1, or GFAP and UCH-L1 are elevated in the sample, and communicating the determination on or from an instrument. The methods may be used to aid in the diagnosis and evaluation of a subject (e.g., a human subject) that has sustained or may have sustained an injury to the head, such as to determine whether the subject is suffering from a mild, moderate, severe, or moderate to severe traumatic brain injury (TBI).

Description

Claims

1. A method comprising the steps of: a. performing at least one assay for ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) and at least one assay for glial fibrillary acidic protein (GFAP) in at least one sample obtained from a human subject, wherein the sample is obtained from the subject within about 48 hours after an actual or suspected injury to the head;b. determining that: 1. the subject’s levels of GFAP and UCH-L1 are elevated when (i) the level of GFAP in the sample is equal to or above about 35 pg/mL and level of UCH-L1 in the sample is below about 400 pg/mL, cannot be determined or is not reported; (ii) the level of GFAP in the sample is equal to or above about 35 pg/mL and level of UCH-L1 in the sample is equal to or above about 400 pg/mL; or (iii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is equal to or above about 400 pg/mL;2. the subject’s levels of GFAP and UCH-L1 are not elevated when the level of GFAP in the sample is below about 35 pg/mL and level of UCH-L1 in the sample is below about 400 pg/mL; or3. the assays for UCH-L1 and GFAP should be repeated when (i) the level of GFAP in the sample is below about 35 pg/mL and the level of UCH-L1 in the sample cannot be determined or is not reported; (ii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is below about 400 pg/mL; or (iii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample cannot be determined or is not reported, andc. communicating the determination from step b (1) - (3) on or from at least one instrument, wherein the instrument is a non-point-of-care device.

2. The method of claim 1, wherein the method further comprises: a. performing a head computed tomography (CT) scan, magnetic resonance imaging (MRI) procedure, or both a CT scan or a MRI procedure on the subject when the subject’s levels of GFAP and UCH-L1 are elevated, orb. not performing a head CT scan or an MRI procedure on the subject when the subject’s levels of GFAP and UCH-L1 are not elevated.

3. The method of claim 1, further comprising diagnosing the subject as having a traumatic brain injury (TBI) when the level of GFAP is equal to or above about 35 pg/mL and the level of UCH-L1 is equal to or above about 400 pg/mL, regardless of whether a head CT scan is negative for a TBI or whether any head CT scan is performed.

4. The method of claim 1, wherein the method further comprises: (a) treating the subject for a mild, moderate, moderate to severe, or severe TBI when the subject’s levels of GFAP and UCH-L1 are elevated;(b) monitoring the subject where the subject’s levels of GFAP and UCH-L1 are elevated; or(c) a combination of (a) and (b).

5. (canceled)

6. The method of claim 1, wherein the sample is taken within about 5 minutes, within about 10 minutes, within about 12 minutes, within about 15 minutes, within about 20 minutes, within about 30 minutes, within about 60 minutes, within about 90 minutes, within about 2 hours, within about 3 hours, within about 4 hours, within about 5 hours, within about 6 hours, within about 7 hours, within about 8 hours, within about 9 hours, within about 10 hours, within about 11 hours, within about 12 hours, within about 13 hours, within about 14 hours, within about 15 hours, within about 16 hours, within about 17 hours, within about 18 hours, within about 19 hours, within about 20 hours, within about 21 hours, within about 22 hours, within about 23 hours, within about 24 hours, within about 25 hours, within about 26 hours, within about 27 hours, within about 28 hours, within about 29 hours, within about 30 hours, within about 31 hours, within about 32 hours, within about 33 hours, within about 34 hours, within about 35 hours, within about 36 hours, within about 37 hours, within about 38 hours, within about 39 hours, within about 40 hours, within about 41 hours, within about 42 hours, within about 43 hours, within about 44 hours, within about 45 hours, within about 46 hours, within about 47 hours, within about 48 hours after the actual or suspected injury to the head or within about 12 hour to within about 48 hours.

7. (canceled)

8. The method of claim 1, wherein the sample is obtained: (a) after the subject sustained an injury to the head caused by physical shaking, blunt impact by an external mechanical or other force that results in a closed or open head trauma, one or more falls, explosions or blasts or other types of blunt force trauma;(b) after the subject has ingested or been exposed to a chemical, toxin or combination of a chemical and toxin; or(c) from a subject that suffers from an autoimmune disease, a metabolic disorder, a brain tumor, hypoxia, a viral infection, a fungal infection, a bacterial infection, meningitis, hydrocephalus, or any combinations thereof.

9. (canceled)

10. (canceled)

11. (canceled)

12. The method of claim 1, wherein the assay is: (a) an immunoassay or a clinical chemistry assay; or (b) a single molecule detection assay.

13. (canceled)

14. The method of claim 1, wherein the amount of the at least one sample is: (a) about 10 µL to about 30 µL; or (b) about 20 µL.

15. (canceled)

16. The method of claim 1, wherein the at least one assay for UCH-L1, at least one assay for GFAP, or at least one assay for UCH-L1 and at least one assay for GFAP is performed in: (a) about 10 to about 20 minutes; or (b) about 15 minutes.

17. (canceled)

18. (canceled)

19. The method of claim 1, wherein the sample is selected from the group consisting of a whole blood sample, a serum sample, and a plasma sample.

20. A system comprising: an assay for ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) and an assay for glial fibrillary acidic protein (GFAP); anda non-point-of-care device for performing the assay for UCH-L1 and the assay for GFAP, whereinthe device determines an amount of UCH-L1 and GFAP in a sample obtained from a subject, andthe amount of UCH-L1 and GFAP determined in the sample are communicated on or from the device as: a. elevated when (i) the level of GFAP in the sample is equal to or above about 35 pg/mL and level of UCH-L1 in the sample is below about 400 pg/mL, cannot be determined or is not reported; (ii) the level of GFAP in the sample is equal to or above about 35 pg/mL and level of UCH-L1 in the sample is equal to or above about 400 pg/mL; or (iii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is equal to or above about 400 pg/mL;b. not elevated when the level of GFAP in the sample is below about 35 pg/mL and level of UCH-L1 in the sample is below about 400 pg/mL; orc. requiring the assays for UCH-L1 and GFAP to be repeated when (i) the level of GFAP in the sample is below about 35 pg/mL and the level of UCH-L1 in the sample cannot be determined or is not reported; (ii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is below about 400 pg/mL; or (iii) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample cannot be determined or is not reported.

21. The system of claim 20, wherein the sample is taken within about 5 minutes, within about 10 minutes, within about 12 minutes, within about 15 minutes, within about 20 minutes, within about 30 minutes, within about 60 minutes, within about 90 minutes, within about 2 hours, within about 3 hours, within about 4 hours, within about 5 hours, within about 6 hours, within about 7 hours, within about 8 hours, within about 9 hours, within about 10 hours, within about 11 hours, within about 12 hours, within about 13 hours, within about 14 hours, within about 15 hours, within about 16 hours, within about 17 hours, within about 18 hours, within about 19 hours, within about 20 hours, within about 21 hours, within about 22 hours, within about 23 hours, within about 24 hours, within about 25 hours, within about 26 hours, within about 27 hours, within about 28 hours, within about 29 hours, within about 30 hours, within about 31 hours, within about 32 hours, within about 33 hours, within about 34 hours, within about 35 hours, within about 36 hours, within about 37 hours, within about 38 hours, within about 39 hours, within about 40 hours, within about 41 hours, within about 42 hours, within about 43 hours, within about 44 hours, within about 45 hours, within about 46 hours, within about 47 hours, within about 48 hours, or within about 12 hour to within about 48 hours after an actual or suspected injury to the head.

22. (canceled)

23. The system of claim 20, wherein the sample is obtained: (a) after the subject sustained an injury to the head caused by physical shaking, blunt impact by an external mechanical or other force that results in a closed or open head trauma, one or more falls, explosions or blasts or other types of blunt force trauma;(b) after the subject has ingested or been exposed to a chemical, toxin or combination of a chemical and toxin; or(c) from a subject that suffers from an autoimmune disease, a metabolic disorder, a brain tumor, hypoxia, a viral infection, a fungal infection, a bacterial infection, meningitis, hydrocephalus, or any combinations thereof.

24. (canceled)

25. (canceled)

26. (canceled)

27. The system of claim 20, wherein the assay is: (a) an immunoassay or a clinical chemistry assay; or (b) a single molecule detection assay.

28. (canceled)

29. The system of claim 20, wherein the amount of the at least one sample is: (a) about 10 µL to about 30 µL; or (b) about 20 uL.

30. (canceled)

31. The system of claim 20, wherein the at least one assay for UCH-L1, at least one assay for GFAP, or at least one assay for UCH-L1 and at least one assay for GFAP is performed in: (a) about 10 to about 20 minutes; or (b) about 15 minutes.

32. (canceled)

33. (canceled)

34. The system of claim 20, wherein the sample is selected from the group consisting of a whole blood sample, a serum sample, and a plasma sample.

35. A method comprising the steps of: a. performing at least one assay for ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), at least one assay for glial fibrillary acidic protein (GFAP), or at least one assay for UCH-L1 and at least one assay for GFAP in at least one sample obtained from a human subject, wherein the sample is obtained from the subject within about 48 hours after an actual or suspected injury to the head;b. determining that: 1. the subject’s levels of GFAP, UCH-L1, or GFAP and UCH-L1 are elevated when (i) the level of GFAP alone in the sample is equal to or above about 35 pg/mL; (ii) the level of GFAP in the sample is equal to or above about 35 pg/mL and level of UCH-L1 in the sample is below about 400 pg/mL, cannot be determined or is not reported; (iii) the level of GFAP in the sample is equal to or above about 35 pg/mL and level of UCH-L1 in the sample is equal to or above about 400 pg/mL; (iv) the level of UCH-L1 alone in the sample is equal to or above 400 pg/mL; or (v) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is equal to or above about 400 pg/mL;2. the subject’s levels of (i) GFAP are not elevated when GFAP alone in the sample is below about 35 pg/mL; (ii) UCH-L1 are not elevated when UCH-L1 alone in the sample is below about 400 pg/mL; or (iii) GFAP and UCH-L1 are not elevated when the level of GFAP in the sample is below about 35 pg/mL and level of UCH-L1 in the sample is below about 400 pg/mL; or3. the assays for UCH-L1 and GFAP should be repeated when (i) the level of UCH-L1 alone in the sample cannot be determined or is not reported; (ii) the level of GFAP in the sample is below about 35 pg/mL and the level of UCH-L1 in the sample cannot be determined or is not reported; (iii) the level of GFAP alone in the sample cannot be determined or is not reported; (iv) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is below about 400 pg/mL; or (v) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample cannot be determined or is not reported, andc. communicating the determination from step b (1) - (3) on or from at least one instrument, wherein the instrument is a non-point-of-care device.

36. The method of claim 35, wherein the method further comprises: a. performing a head computed tomography (CT) scan, magnetic resonance imaging (MRI) procedure, or both a CT scan or a MRI procedure on the subject when the subject’s levels of GFAP, UCH-L1, or GFAP and UCH-L1 are elevated, orb. not performing a head CT scan or an MRI procedure on the subject when the subject’s levels of GFAP, UCH-L1, or GFAP and UCH-L1 are not elevated.

37. The method of claim 35, further comprising diagnosing the subject as having a traumatic brain injury (TBI) when the level of GFAP is equal to or above about 35 pg/mL, the level of UCH-L1 is equal to or above about 400 pg/mL, or level of GFAP is equal to or above about 35 pg/mL and the level of UCH-L1 is equal to or above about 400 pg/mL, regardless of whether a head CT scan is negative for a TBI or whether any head CT scan is performed.

38. The method of claim 35, wherein the method further comprises (a) treating the subject for a mild, moderate, moderate to severe, or severe TBI when the subject’s levels of GFAP and UCH-L1 are elevated;(b) monitoring the subject where the subject’s levels of GFAP and UCH-L1 are elevated; or(c) a combination of (a) and (b).

39. (canceled)

40. The method of claim 35, wherein the sample is taken within about 5 minutes, within about 10 minutes, within about 12 minutes, within about 15 minutes, within about 20 minutes, within about 30 minutes, within about 60 minutes, within about 90 minutes, within about 2 hours, within about 3 hours, within about 4 hours, within about 5 hours, within about 6 hours, within about 7 hours, within about 8 hours, within about 9 hours, within about 10 hours, within about 11 hours, within about 12 hours, within about 13 hours, within about 14 hours, within about 15 hours, within about 16 hours, within about 17 hours, within about 18 hours, within about 19 hours, within about 20 hours, within about 21 hours, within about 22 hours, within about 23 hours, within about 24 hours, within about 25 hours, within about 26 hours, within about 27 hours, within about 28 hours, within about 29 hours, within about 30 hours, within about 31 hours, within about 32 hours, within about 33 hours, within about 34 hours, within about 35 hours, within about 36 hours, within about 37 hours, within about 38 hours, within about 39 hours, within about 40 hours, within about 41 hours, within about 42 hours, within about 43 hours, within about 44 hours, within about 45 hours, within about 46 hours, within about 47 hours, within about 48 hours, or within about 12 hour to within about 48 hours after the actual or suspected injury to the head.

41. (canceled)

42. The method of claim 35, wherein the sample is obtained: (a) after the subject sustained an injury to the head caused by physical shaking, blunt impact by an external mechanical or other force that results in a closed or open head trauma, one or more falls, explosions or blasts or other types of blunt force trauma;(b) after the subject has ingested or been exposed to a chemical, toxin or combination of a chemical and toxin; or(c) from a subject that suffers from an autoimmune disease, a metabolic disorder, a brain tumor, hypoxia, a viral infection, a fungal infection, a bacterial infection, meningitis, hydrocephalus, or any combinations thereof.

43. (canceled)

44. (canceled)

45. (canceled)

46. The method of claim 35, wherein the assay is: (a) an immunoassay or a clinical chemistry assay; or (b) a single molecule detection assay.

47. (canceled)

48. The method of claim 35, wherein the amount of the at least one sample is: (a) about 10 µL to about 30 µL; or (b) about 20 µL..

49. (canceled)

50. The method of claim 35, wherein the at least one assay for UCH-L1, at least one assay for GFAP, or at least one assay for UCH-L1 and at least one assay for GFAP is performed in: (a) about 10 to about 20 minutes; or (b) about 15 minutes.

51. (canceled)

52. The method of claim 35, wherein the subject has sustained an orthopedic injury and an actual or suspected injury to the head.

53. The method of claim 35, wherein the sample is selected from the group consisting of a whole blood sample, a serum sample, and a plasma sample.

54. A system comprising: an assay for ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), an assay for glial fibrillary acidic protein (GFAP), or an assay for UCH-L1 and an assay for GFAP; anda non-point-of-care device for performing the assay for UCH-L1, the assay for GFAP, or the assay for UCH-L1 and the assay for GFAP, whereinthe device determines an amount of UCH-L1, GFAP, or UCH-L1 and GFAP in a sample obtained from a subject, andthe amount of UCH-L1, GFAP, or UCH-L1 and GFAP determined in the sample are communicated on or from the device as: a. elevated when (i) the level of GFAP alone in the sample is equal to or above about 35 pg/mL; (ii) the level of GFAP in the sample is equal to or above about 35 pg/mL and level of UCH-L1 in the sample is below about 400 pg/mL, cannot be determined or is not reported; (iii) the level of GFAP in the sample is equal to or above about 35 pg/mL and level of UCH-L1 in the sample is equal to or above about 400 pg/mL; (iv) the level of UCH-L1 alone in the sample is equal to or above about 400 pg/mL; or (v) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is equal to or above about 400 pg/mL;b. not elevated when (i) the level of GFAP alone in the sample is below about 35 pg/mL; (ii) the level of UCH-L1 alone in the sample is below about 400 pg/mL; or (iii) the level of GFAP in the sample is below about 35 pg/mL and level of UCH-L1 in the sample is below about 400 pg/mL; orc. requiring the assays for UCH-L1 and GFAP to be repeated when (i) the level of UCH-L1 alone in the sample cannot be determined or is not reported; (ii) the level of GFAP in the sample is below about 35 pg/mL and the level of UCH-L1 in the sample cannot be determined or is not reported; (iii) the level of GFAP alone in the sample cannot be determined or is not reported; (iv) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample is below about 400 pg/mL; or (v) the level of GFAP in the sample cannot be determined or is not reported and the level of UCH-L1 in the sample cannot be determined or is not reported.

55. The system of claim 54, wherein the sample is taken within about 5 minutes, within about 10 minutes, within about 12 minutes, within about 15 minutes, within about 20 minutes, within about 30 minutes, within about 60 minutes, within about 90 minutes, within about 2 hours, within about 3 hours, within about 4 hours, within about 5 hours, within about 6 hours, within about 7 hours, within about 8 hours, within about 9 hours, within about 10 hours, within about 11 hours, within about 12 hours, within about 13 hours, within about 14 hours, within about 15 hours, within about 16 hours, within about 17 hours, within about 18 hours, within about 19 hours, within about 20 hours, within about 21 hours, within about 22 hours, within about 23 hours, within about 24 hours, within about 25 hours, within about 26 hours, within about 27 hours, within about 28 hours, within about 29 hours, within about 30 hours, within about 31 hours, within about 32 hours, within about 33 hours, within about 34 hours, within about 35 hours, within about 36 hours, within about 37 hours, within about 38 hours, within about 39 hours, within about 40 hours, within about 41 hours, within about 42 hours, within about 43 hours, within about 44 hours, within about 45 hours, within about 46 hours, within about 47 hours, within about 48 hours, or within about 12 hour to within about 48 hours after an actual or suspected injury to the head.

56. (canceled)

57. The system of claim 54, wherein the sample is obtained after: (a) the subject sustained an injury to the head caused by physical shaking, blunt impact by an external mechanical or other force that results in a closed or open head trauma, one or more falls, explosions or blasts or other types of blunt force trauma;(b) after the subject has ingested or been exposed to a chemical, toxin or combination of a chemical and toxin; or(c) from a subject that suffers from an autoimmune disease, a metabolic disorder, a brain tumor, hypoxia, a viral infection, a fungal infection, a bacterial infection, meningitis, hydrocephalus, or any combinations thereof.

58. (canceled)

59. (canceled)

60. (canceled)

61. The system of claim 54, wherein the assay is: (a) an immunoassay or a clinical chemistry assay; or (b) a single molecule detection assay.

62. (canceled)

63. The system of claim 54, wherein the amount of the at least one sample: (a) about 10 µL to about 30 µL; or (b) about 20 µL.

64. (canceled)

65. The system of claim 54, wherein the at least one assay for UCH-L1, at least one assay for GFAP, or at least one assay for UCH-L1 and at least one assay for GFAP is performed in: (a) about 10 to about 20 minutes; or (b) about 15 minutes.

66. (canceled)

67. (canceled)

68. The system of claim 54, wherein the sample is selected from the group consisting of a whole blood sample, a serum sample, and a plasma sample.