Advanced Detection of Sepsis

• US 20110118569A1
• Filed: 04/02/2009
• Published: 05/19/2011
• Est. Priority Date: 04/03/2008
• Status: Active Grant

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Garrett, James, Shi, Song, Moore, Richard L.
• Granted Patent: US 8,669,113 B2
• Time in Patent Office: Days
• US Class Current: 600/309
• CPC Class Codes: C12Q 1/6881 for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 2333/90209 acting on NADH or NADPH (1.6), e.g. those with a heme protein as acceptor (1.6.2) (general), Cytochrome-b5 reductase (1.6.2.2) or NADPH-cytochrome P450 reductase (1.6.2.4)
• CPC Class Codes: G01N 2333/91051 Acyltransferases other than aminoacyltransferases (general) (2.3.1)
• CPC Class Codes: G01N 2800/26 Infectious diseases, e.g. generalised sepsis
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743

Abstract

The present invention relates to methods, monitors and systems measuring lysophosphatidylcholine, its derivatives and/or procalcitonin as well as at least one clinical marker (e.g. temperature or respiration rate) and/or at least one biomarker for the early detection of sepsis in a subject.

Claims

1. A method for the advanced detection of sepsis in a subject, comprising the steps of:

(a) measuring at a plurality of time points an amount of lysophosphatidylcholine in fluid or tissue of the subject; and

(b) measuring at a plurality of time points one or more clinical markers of the subject,

to detect sepsis in the subject.

2. The method of claim 1, wherein one clinical marker is measured.

3. The method of claim 1, wherein respiratory rate is measured.

4. The method of claim 1, wherein two or more clinical markers are measured.

5. The method of claim 1, wherein respiratory rate and temperature are measured.

6. The method of claim 1, wherein said lysophosphatidylcholine comprises 1-O-palmitoyl-2-lyso-sn-glycero-3-phosphocholine and 1-O-stearoyl-2-lyso-sn-glycero-3-phosphocholine.

7. The method of claim 1, wherein said advanced detection is at least 12 hours prior to the onset of sepsis.

8. The method of claim 1, wherein the amount is measured 12, 24, 36 or 48 hours prior to the onset of sepsis.

9. The method of claim 1, wherein decreasing amounts of lysophosphatidylcholine detect sepsis.

10. The method of claim 1, wherein free lysophosphatidylcholine is measured.

11. The method of claim 1, wherein bound lysophosphatidylcholine is measured.

12. The method of claim 1, wherein total lysophosphotatidylcholine is measured.

30. The method of claim 1 or 13, further comprising measuring at a plurality of time points an amount of one or more biomarkers in fluid or tissue of the subject.

31. The method of claim 30, wherein a difference in the amount of one or more biomarkers measured at a plurality of time points detects sepsis.

32. The method of claim 30, wherein the one or more biomarkers are selected from the group consisting of endotoxin, bacterial DNA, protein C, protein S, procalcitonin, C-reactive protein, LBP LPS-binding protein, fibrin degrading products, HLA-DR, cell surface proteins CD-14 and CD-64, E-selectin, cortisol, ACTH, surface-bound tumor necrosis factor receptor I, surface-bound tumor necrosis factor receptor II, tumor necrosis factor alpha, interleukins IL-6, IL-8 and IL-10, D-dimer, prothrombin, antithrombin III, activated partial thromboplastin, plasminogen activator inhibitor-1, soluble thrombomodulin, thrombin activatable fibrinolysis inhibitor, copeptin, high mobility group box 1, triggering receptor expressed on myeloid cells 1, and albumin.

33. The method of claim 32 wherein the one or more biomarkers comprise procalcitonin.

44. The method of any one of claim 1, 13, or 34, wherein the subject is SIRS-negative.

45. The method of any one of claim 1, 13, or 34, wherein the subject is SIRS-positive.

46. The method of any one of claim 1, 13, or 34, wherein the amount is measured by spectrometry, chromatography, immunoassay, electrophoresis or enzymatic assay.

47. The method of any one of claim 1, 13, or 34, wherein the fluid or tissue is blood, plasma, saliva, serum, sputum, urine, cells, cellular extract or tissue biopsy.

13. A method for the advanced detection of sepsis in a subject, comprising the steps of:

(a) measuring at a plurality of time points an amount of a compound according to formula (I)

14. The method of claim 13, wherein one clinical marker is measured.

15. The method of claim 13, wherein respiratory rate is measured.

16. The method of claim 13, wherein two or more clinical markers are measured.

17. The method of claim 13, wherein respiratory rate and temperature are measured.

18. The method of claim 13, wherein R is saturated acyl.

19. The method of claim 13, wherein R is C₁₄-C₂₂ acyl.

20. The method of claim 13, wherein R is C₁₆-C₂₀ acyl.

21. The method of claim 13, wherein R is C₁₆-C₁₈ acyl.

22. The method of claim 13, wherein R is C₁₆ saturated acyl.

23. The method of claim 13, wherein R is C₁₈ saturated acyl.

24. The method of claim 13, wherein R is unbranched C₁₆-C₁₈ acyl.

25. The method of claim 13, wherein R is palmitoyl.

26. The method of claim 13, wherein R is stearoyl.

27. The method of claim 13, wherein said advanced detection is at least 12 hours prior to the onset of sepsis.

28. The method of claim 13, wherein the amount is measured 12, 24, 36 or 48 hours prior to the onset of sepsis.

29. The method of claim 13, wherein the difference between the amounts measured for the compound correlates inversely with detection of sepsis.

34. A method for the advanced detection of sepsis in a subject, comprising the steps of:

(a) measuring at a plurality of time points an amount of procalcitonin in fluid or tissue of the subject; and

(b) measuring at a plurality of time points one or more clinical markers of the subject,

to detect sepsis in the subject.

35. The method of claim 34, wherein the one or more clinical marker(s) are selected from the group consisting of respiratory rate, temperature, heart rate, systolic blood pressure, diastolic blood pressure, mean artery pressure, white blood cell count, monocyte count, lymphocyte count, granulocyte count, neutrophil count, immature neutrophil to total neutrophil ratio, platelet count, serum creatinine concentration, urea concentration, lactate concentration, glucose concentration, base excess, pO₂ and HCO₃⁻

concentration.

36. The method of claim 34, wherein one clinical marker is measured.

37. The method of claim 36, wherein respiratory rate is measured.

38. The method of claim 34, wherein two or more clinical markers are measured.

39. The method of claim 38, wherein respiratory rate and temperature are measured.

40. The method of claim 34, wherein the advanced detection is at least 12 hours prior to the onset of sepsis.

41. The method of claim 34, wherein a body temperature greater than 38°

C. detects sepsis.

42. The method of claim 34, wherein a body temperature less than 36°

C. detects sepsis.

43. The method of claim 34, wherein a respiratory rate greater than 20 breaths per minute detects sepsis.

48. A monitor for the advanced detection of sepsis in a subject comprising:

(a) a sensor module capable of measuring at a plurality of time points one or more clinical markers of the subject; and

(b) a chemistry module capable of measuring at a plurality of time points an amount of lysophosphatidylcholine in fluid or tissue of the subject.

49. The monitor of claim 48 comprising one or more sensor modules capable of measuring two or more clinical markers.

50. The monitor of claim 49 capable of measuring respiratory rate and temperature.

52. The monitor of claim 48 or 51, wherein the one or more sensor modules are capable of continuous measuring.

60. The monitor of claim 48 or 51, wherein the chemistry module is capable of contacting a sample from the fluid or tissue of the subject with:

an enzyme or reagent capable of reacting lysophosphatidylcholine to form glycerophosphatidylcholine;

an enzyme or reagent capable of reacting glycerophosphatidylcholine to form choline;

an enzyme or reagent capable of reacting choline, water and oxygen to form peroxide;

a peroxidase and

a fluorogenic substrate of said peroxidase

under conditions suitable for formation of a fluorescent product wherein the fluorescent product indicates lysophosphatidylcholine.

61. The monitor of claim 60 wherein said enzyme or reagent capable of reacting lysophosphatidylcholine to form glycerophosphatidylcholine is a lysophospholipase.

62. The monitor of claim 60 wherein said enzyme or reagent capable of converting lysophosphatidylcholine to glycerophosphatidylcholine is EC 3.1.1.5.

63. The monitor of claim 60 wherein said enzyme or reagent capable of reacting glycerophosphatidylcholine to form choline is a glycerophosphatidylcholine diesterase.

64. The monitor of claim 60 wherein said enzyme or reagent capable of reacting glycerophosphatidylcholine to form choline is EC 3.1.4.2.

65. The monitor of claim 60 wherein said enzyme or reagent capable of reacting choline to form peroxide is a choline oxidase.

66. The monitor of claim 60 wherein said enzyme or reagent capable of reacting choline to peroxide is EC 1.1.3.17.

67. The monitor of claim 60 wherein said peroxidase is horseradish peroxidase.

68. The monitor of claim 60 wherein said fluorogenic substrate is 10-acetyl-3,7-dihydroxyphenoxazine and said fluorescent product is 7-hydroxy-3H-phenoxazin-3-one.

69. The monitor of claim 60 wherein said fluorescent product is detected by fluorescence detection.

70. The monitor of claim 60 wherein said enzyme or reagent capable of contacting the sample is a lysophospholipase, a glycerophosphatidylcholine diesterase, a choline oxidase, or a peroxidase;

and wherein said fluorogenic substrate capable of forming a fluorescent product is 10-acetyl-3,7-dihydroxyphenoxazine.

71. The monitor of claim 60 wherein the monitor is capable of contacting said sample with EC 3.1.1.5, EC 3.1.4.2, EC 1.1.3.17, horseradish peroxidase and 10-acetyl-3,7-dihydroxyphenoxazine under conditions suitable for formation of 7-hydroxy-3H-phenoxazin-3-one wherein 7-hydroxy-3H-phenoxazin-3-one indicates lysophosphatidylcholine.

72. The monitor of claim 60 wherein the amount of fluorescent product is capable of indicating the amount of lysophosphatidylcholine.

73. A system for the advanced detection of sepsis in a subject comprising:

(a) the monitor of claim 48;

(b) a computational device capable of combining the clinical marker and lysophosphatidylcholine measurements into a result, comprising;

(i) a device capable of receiving the clinical marker and lysophosphatidylcholine measurements from the monitor;

(ii) a microprocessor with an algorithm capable of combining the measurements into a result; and

(iii) a device capable of transmitting the result to a module capable of storing, displaying and/or transmitting; and

(c) a module capable of storing, displaying or transmitting the result.

74. The system of claim 73 wherein the device in (i) and the microprocessor in (ii) are the same device.

81. The system of any one of claims 73-80, wherein the algorithm is selected from discriminant analysis, quadratic discriminant analysis, logical regression analysis, regression classifiers, neural networks, and combinations thereof.

82. The system of any one of claims 73-80, wherein the computational device is further capable of comparing the lysophosphatidylcholine amount to a reference amount indicative of the amounts of lysophosphatidylcholine in fluids or tissues of a plurality of individuals that have, or will have, sepsis.

51. A monitor for the advanced detection of sepsis in a subject comprising:

(a) a first sensor module capable of measuring at a plurality of time points the subject'"'"'s temperature;

(b) a second sensor module capable of measuring at a plurality of time points the subject'"'"'s respiratory rate; and

(c) a chemistry module capable of measuring at a plurality of time points an amount of lysophosphatidylcholine in fluid or tissue of the subject.

53. The monitor of claim 51, wherein the first sensor module is capable of connecting to a temperature probe in the subject.

54. The monitor of claim 53, wherein the temperature probe is capable of being applied externally to the subject'"'"'s skin.

55. The monitor of claim 53, wherein the temperature probe is capable of being applied internally to the subject.

56. The monitor of claim 55, wherein the temperature probe is capable of being applied to the subject'"'"'s esophagus or rectum.

57. The monitor of claim 51, wherein the second sensor module is capable of connecting to a motion sensor.

58. The monitor of claim 57, wherein the motion sensor is a flow meter.

59. The monitor of claim 58, wherein the flow meter is in a respirator.

77. A system for the advanced detection of sepsis in a subject comprising:

(a) the monitor of claim 51;

(b) a computational device capable of combining the temperature, respiratory rate, and lysophosphatidylcholine measurements into a result, comprising;

(i) a device capable of receiving the temperature, respiratory rate, and lysophosphatidylcholine measurements from the monitor;

(ii) a microprocessor with an algorithm capable of combining the measurements into a result; and

(iii) a device capable of transmitting the result to a module capable of storing, displaying and/or transmitting; and

(c) a module capable of storing, displaying or transmitting the result.

78. The system of claim 77 wherein the device in (i) and the microprocessor in (ii) are the same device.

75. A system for the advanced detection of sepsis in a subject comprising:

(a) a sensor module capable of measuring at a plurality of time points one or more clinical markers of the subject;

(b) a chemistry module capable of measuring at a plurality of time points an amount of lysophosphatidylcholine in fluid or tissue of the subject; and

(c) a computational device capable of combining the clinical marker and lysophosphatidylcholine measurements into a result, comprising;

(i) a device capable of receiving the clinical marker and lysophosphatidylcholine measurements from the modules;

(ii) a microprocessor with an algorithm capable of combining the measurements into a result; and

(iii) a device capable of transmitting the result to a module capable of storing, displaying and/or transmitting; and

(d) a module capable of storing, displaying or transmitting the result.

76. The system of claim 75 wherein the device in (i) and the microprocessor in (ii) are the same device.

79. A system for the advanced detection of sepsis in a subject comprising:

(a) a first sensor module capable of measuring at a plurality of time points the subject'"'"'s temperature;

(b) a second sensor module capable of measuring at a plurality of time points the subject'"'"'s respiratory rate;

(c) a chemistry module capable of measuring at a plurality of time points an amount of lysophosphatidylcholine in fluid or tissue of the subject; and

(d) a computational device capable of combining the temperature, respiratory rate, and lysophosphatidylcholine measurements into a result, comprising;

(i) a device capable of receiving the temperature, respiratory rate, and lysophosphatidylcholine measurements from the modules;

(ii) a microprocessor with an algorithm capable of combining the measurements into a result; and

(iii) a device capable of transmitting the result to a module capable of storing, displaying and/or transmitting; and

(e) a module capable of storing, displaying or transmitting the result.

80. The system of claim 79 wherein the device in (i) and the microprocessor in (ii) are the same device.

83. A system for the advanced detection of sepsis in a subject comprising:

(a) a sensor module capable of measuring at a plurality of time points one or more clinical markers of the subject;

(b) a chemistry module capable of measuring at a plurality of time points an amount of lysophosphatidylcholine or procalcitonin in fluid or tissue of the subject; and

(c) a computational device capable of combining the measurements in (a) and the measurements in (b) into a result, comprising;

(i) a device capable of receiving the measurements in (a) and the measurements in (b) from the modules;

(ii) a microprocessor with an algorithm capable of combining the measurements into a result; and

(iii) a device capable of transmitting the result to a module capable of storing, displaying and/or transmitting; and

(d) a module capable of storing, displaying or transmitting the result.

84. The system of claim 83 wherein the device in (i) and the microprocessor in (ii) are the same device.