Biomarker-based methods and biochips for aiding the diagnosis of stroke

Abstract

The present invention provides biomarker-based methods for diagnosing stroke in a patient suspected of having suffered a stroke, and also for discriminating between ischemic stroke and transient ischemic attack. Substrates comprising probes for specific combinations of biomarkers useful in the methods of the invention are also described.

1 .- 25 . (canceled) 
     
     
         26 . A method of aiding the diagnosis of ischaemic stroke in a patient suspected of having a stroke, comprising
 i) determining the concentration of VCAM-1 and one or more biomarkers selected from h-FABP, IL-6 and CRP in an in vitro sample obtained from the patient; and   ii) establishing the significance of the concentration of the biomarkers by comparing the concentration value for each biomarker with a corresponding control value, wherein the corresponding control value is the concentration value for the corresponding biomarker determined from an in vitro sample obtained from a transient ischaemic attack patient or patients.   
     
     
         27 . A method according to  claim 26 , wherein the method is used to differentially diagnose between ischemic stroke and a transient ischaemic attack. 
     
     
         28 . A method according to  claim 26 , wherein each of the patient and control biomarker concentration values is inputted into a statistical algorithm or algorithms to produce an output value that indicates whether ischaemic stroke has occurred. 
     
     
         29 . A method according to  claim 28 , wherein the statistical algorithm includes a logistic regression equation. 
     
     
         30 . A substrate comprising probes for VCAM-1 and at least one other biomarker selected from h-FABP, IL-6 and CRP for use in a method for aiding the diagnosis of ischaemic stroke in a patient according to  claim 26 . 
     
     
         31 . A substrate according to  claim 30 , wherein the substrate has the probes immobilised thereon. 
     
     
         32 . A substrate according to  claim 30 , wherein the substrate is a biochip. 
     
     
         33 . Use of a substrate comprising probes for VCAM-1 and at least one other biomarker selected from h-FABP, IL-6 and CRP in a method for aiding the diagnosis of ischaemic stroke in a patient according to  claim 26 . 
     
     
         34 . Use of VCAM-1 and one or more of h-FABP, IL-6 and CRP, as biomarkers of ischemic stroke and/or as differentiators between ischemic stroke and a transient ischaemic attack. 
     
     
         35 . A method for diagnosing stroke in a patient suspected of having a stroke, comprising determining the concentration of at least two biomarkers in an in vitro sample obtained from the patient and establishing the significance of the concentration of the biomarkers by comparing the concentration value for each biomarker with a corresponding control value, wherein the at least two biomarkers are selected from ICAM-1, L-selectin, P-selectin, VCAM-1, IL-6, sTNFR1, D-dimer and CRP, and wherein at least one of the two biomarkers is selected from ICAM-1, L-selectin, P-selectin and VCAM-1. 
     
     
         36 . A method according to  claim 35 , wherein the at least two biomarkers are selected from (i) ICAM-1 or VCAM-1 and (ii) L-selectin or P-select in. 
     
     
         37 . A method according to  claim 35 , wherein the at least two biomarkers are ICAM-1 and L-selectin. 
     
     
         38 . A method according to  claim 36 , further comprising determining the sample concentration of one or more biomarkers selected from IL-6, sTNFR1, D-dimer and CRP. 
     
     
         39 . A method according to  claim 35 , further comprising determining the sample concentration of h-FABP. 
     
     
         40 . A method according to  claim 26 , wherein each of the patient and control biomarker concentration values is inputted into a statistical algorithm or algorithms to produce an output value that indicates whether a stroke has occurred. 
     
     
         41 . A method according to  claim 40 , wherein the statistical algorithm includes a logistic regression equation. 
     
     
         42 . A method according to  claim 35 , wherein the combination of two or more biomarkers corresponds to any of the biomarker combinations listed in Tables 1 and 2. 
     
     
         43 . A method according to  claim 35 , wherein if stoke is diagnosed, a second method is carried out in order to differentially diagnose the stroke type, the second method comprising:
 i) determining the concentration of VCAM-1 and one or more biomarkers selected from h-FABP, IL-6 and CRP in an in vitro sample obtained from the patient; and   ii) establishing the significance of the concentration of the biomarkers by comparing the concentration value for each biomarker with a corresponding control value, wherein the corresponding control value is the concentration value for the corresponding biomarker determined from an in vitro sample obtained from a transient ischaemic attack patient or patients.   
     
     
         44 . A substrate comprising probes for at least two biomarkers selected from ICAM-1, L-selectin, P-selectin, VCAM-1, IL-6, sTNFR1, D-dimer and CRP for use in a method for diagnosing stroke in a patient according to  claim 35 , wherein the substrate comprises a probe for at least one of ICAM-1, L-selectin, P-selectin and VCAM-1. 
     
     
         45 . A substrate according to  claim 44 , further comprising a probe for h-FABP. 
     
     
         46 . A substrate according to  claim 44 , wherein the substrate has at least two probes immobilised thereon. 
     
     
         47 . A substrate according to  claim 44 , wherein the substrate is a biochip. 
     
     
         48 . Use of a substrate comprising probes for at least two biomarkers selected from ICAM-1, L-selectin, P-selectin, VCAM-1, IL-6, sTNFR1, D-dimer and CRP in a method for diagnosing stroke in a patient according to  claim 35 , wherein the substrate comprises a probe for at least one of ICAM-1, L-selectin, P-selectin and VCAM-1. 
     
     
         49 . Use according to  claim 48 , wherein the substrate further comprises a probe for h-FABP. 
     
     
         50 . A method according to  claim 26 , wherein the sample is a blood, serum or plasma sample.