Highly Sensitive Biomarker Panels

Abstract

Cardiovascular disease, e.g., congestive heart failure, is often first diagnosed after the onset of clinical symptoms, eliminating potential for early intervention. The invention provides a multi-marker immunoassay, including cardiac pathology and vascular inflammation biomarkers, yielding a more sensitive assay for early detection of CHF in plasma. A panel consisting of cardiac pathology (cTnI, BNP) and vascular inflammation (IL-6, TNFα, IL-17a) biomarkers provided a sensitivity of 94% for association with CHF.

1 - 21 . (canceled) 
     
     
         22 . A method for detecting or monitoring a cardiovascular condition associated with cardiomyocyte damage in a subject, comprising
 detecting the concentration of two or more biomarkers in a blood, serum or plasma sample from the subject, wherein the biomarkers comprise Troponin-I (cTnI) and Interleukin 17a (IL-17a), wherein the cTnI is detected in a single molecule counting assay having a limit of detection for cTnI of less than 3 pg/mL and a coefficient of variation of less than 10%, wherein the cTnI assay comprises contacting the sample with an antibody specific for cTnI and determining the amount of specific binding between the antibody and the cTnI in the sample,   comparing the concentration of the biomarkers to threshold concentrations for the biomarkers, and   determining the presence of the cardiovascular condition when the concentration of the biomarkers in the sample is greater than threshold concentrations for the biomarkers.   
     
     
         23 . The method of  claim 22 , wherein the antibody comprises a label comprising a fluorescent moiety that is detected in an interrogation space defined by an electromagnetic radiation source, wherein a single molecule of the antibody comprising the label corresponds to a single molecule of the cTnI. 
     
     
         24 . The method of  claim 22 , wherein the two or more biomarkers comprise at least one of Interleukin 6 (IL-6), B-type Natriuretic Peptide (BNP), proBNP and NT-proBNP, and Tumor Necrosis Factor alpha (TNF-α). 
     
     
         25 . The method of  claim 22 , wherein the cardiovascular condition associated with cardiomyocyte damage is selected from the group consisting of congestive heart failure (CHF), atherosclerosis, angina pectoris, atrial fibrillation, myocardial ischemia, myocardial infarction, myocarditis, hypertrophic cardiomyopathy and cholesterol embolism. 
     
     
         26 . A method for detecting congestive heart failure in a subject, comprising detecting whether the level of two or more members of a panel of biomarkers in a sample from a subject are elevated compared to the level of the two or more biomarkers in a normal population, wherein the members of the panel comprise Troponin-I (cTnI) and Interleukin 17a (IL-17a), and wherein the cTnI is detected in a single molecule counting assay having a limit of detection for cTnI of less than 3 pg/mL and a coefficient of variation of less than 10%, wherein the cTnI assay comprises contacting the sample with an antibody specific for cTnI and determining the amount of specific binding between the antibody and the cTnI in the sample, 
     
     
         27 . The method of  claim 26 , wherein the antibody comprises a label comprising a fluorescent moiety that is detected in an interrogation space defined by an electromagnetic radiation source, wherein a single molecule of the antibody comprising the label corresponds to a single molecule of the cTnI. 
     
     
         28 . The method of claim  1 , wherein the panel of biomarkers further comprises at least one of Interleukin 6 (IL-6), B-type Natriuretic Peptide (BNP), proBNP and NT-proBNP, and Tumor Necrosis Factor alpha (TNF-α).