Method and means for diagnosing a human sepsis

1 . A method of early detection and diagnosis of a human sepsis, comprising the following steps:

a) providing a blood sample of a patient, wherein said blood sample is a whole blood sample, a platelet-rich plasma sample, a platelet suspension, or a platelet pellet,

b) stimulating a platelet-specific (hem-)immunoreceptor tyrosine-based activation motif ((hem-)ITAM receptor by adding a (hem-)ITAM receptor agonistic agent to said sample, wherein said agonistic agent comprises collagen-related peptide cross-linked (CRP-XL) and/or convulxin,

c) measuring a platelet function level,

wherein said patient has an age of >1 year;

wherein said platelet function level is characterized by at least one parameter selected from the group consisting of a surface presentation of P-selectin, an activated integrin αIIb/β3, a platelet aggregation, a mepacrine uptake/release, and a phosphorylation status of Syk, LAT, SHP-1, SHP-2, FcRγ, and PLCγ2 or a combination thereof;

wherein the platelet function level measured is compared to a reference value and/or a reference sample of a healthy person not suffering from sepsis; and

wherein said method further comprises a step of determining that a patient has the sepsis or is at risk of developing the sepsis, if the platelet function level in said patient is decreased compared to the reference value and/or the reference sample.

2 . The method according to claim 1 , wherein said agonistic agent comprises both CRP-XL and convulxin.

3 . The method according to claim 1 , wherein said method is used for an early diagnosis of the sepsis.

4 . The method according to claim 1 , wherein said measuring is performed by a method selected from flow cytometry, aggregometry, detection and quantification of nucleic acids by polymerase chain reaction (PCR) or real time (quantitative) qPCR, enzyme-linked immunosorbent assay (ELISA), western blot, chromatography, and mepacrine assay.

5 . The method according to claim 4 , wherein said aggregometry is performed by any of light transmission aggregometry, impedance aggregometry, multiple electrode aggregometry, lumino-aggregometry, and microscale aggregometry, wherein a decreased aggregation of platelets is an indicator for sepsis, compared to a reference value and/or a reference sample of a healthy person not suffering from sepsis.

6 . The method according to claim 1 , wherein said measuring is performed by means of:

i) a flow cytometric analysis of the surface presentation of P-selectin, and/or of the activated integrin αIIb/β3, and/or mepacrine uptake/release, and/or

ii) an aggregometry, and/or

iii) a quantification of the phosphorylation status of Syk, LAT, SHP-1, SHP-2, FcRγ, and/or PLCγ2.

7 . The method according to claim 6 , wherein said measuring is performed by means of i), ii), and iii).

8 . The method according to claim 6 ,

wherein said flow cytometric analysis of the surface presentation of P-selectin is performed using an antibody against a P-selectin, and/or

wherein said flow cytometric analysis of the activated integrin αIIb/β3 is performed using an antibody against an activated conformation of integrin αIIb/β3, by fluorophore-conjugated fibrinogen,

wherein a decreased binding of said antibody against P-selectin and/or said antibody against an activated conformation of integrin αIIb/β3 is an indicator for sepsis, compared to a reference value and/or a reference sample of a healthy person not suffering from sepsis.

9 . The method according to claim 8 , wherein the antibody against a P-selectin is an anti-CD62P antibody.

10 . The method according to claim 6 , wherein said quantification of the phosphorylation status is performed by western blot using an antibody against phosphorylated Syk, phosphorylated LAT, phosphorylated SHP-1, phosphorylated SHP-2a, phosphorylated FcRγ, and/or phosphorylated PLCγ2, wherein a hypophosphorylation is an indicator for sepsis, compared to a reference value and/or a reference sample of a healthy person not suffering from sepsis.

11 . The method according to claim 1 , wherein said method is used for monitoring a patient's health condition,

wherein said monitoring comprises performing steps a)-c) at a time point t 1 and a time point t 2 , or

wherein said monitoring comprises performing step a) at a time point t 1 and a time point t 2 , and performing steps b)-c) subsequent to performing said step a) at said time point t 2 ,

wherein said time point t 2 is after said time point t 1 .

12 . The method according to claim 11 , wherein an increase in the platelet function level from said time point t 1 compared to the platelet function level from said time point t 2 indicates an amelioration of the patient's health condition or wherein a decrease in the platelet function level from said time point t 1 compared to the platelet function level from said time point t 2 indicates a worsening of the patient's health condition.

13 . The method according to claim 11 , wherein a time interval between said time point t 1 and said time point t 2 is between 6 h and 7 days.

14 . A kit for diagnosing a human sepsis comprising:

an agonistic agent for stimulating a platelet-specific (hem-)ITAM receptor, said agonistic agent comprising CRP-XL and/or convulxin,

auxiliary compounds for performing the method as defined in claim 1 ,

optionally comprising instructions for comparing a platelet function level of a patient to a reference value and/or a reference sample of a healthy person not suffering from sepsis, wherein a decrease in the platelet function level of the patient compared to the reference value and/or the reference sample indicates a human sepsis.

15 . A method for diagnosing a human sepsis, wherein said method comprises the use of a kit comprising:

an agonistic agent for stimulating a platelet-specific (hem-)ITAM receptor, said agonistic agent comprising CRP-XL and/or convulxin,

auxiliary compounds for performing the method as defined in claim 1 .