ABSTRACT Heart disease is the leading cause of death in the United States, responsible for more than 630,000 deaths annually. Cardiovascular biomarkers play a fundamental role in the diagnosis, risk stratification, and treatment of a wide variety of cardiovascular disorders. Currently there are several testing devices and techniques to quantify the levels of cardiac biomarkers; however, these assays are designed exclusively for clinical settings and are engineered for the detection of individual biomarkers. The objective of this project is to develop an inkjet-printed, plasmonically enhanced, single-step, multiplex point-of-care test for the quantification of key cardiovascular biomarkers, specifically NT-proBNP, cTnT, and CRP. The proposed biomarkers are central to the diagnosis and management of a wide spectrum of cardiovascular conditions, including acute chest pain and myocardial infarction, acute and chronic heart failure, and primary and secondary prevention. The technological innovation of this project stems from the integration of two state-of-the-art platforms—1) a single step, multiplex point-of-care protein microarray with high sensitivity and low background noise and 2) a plasmonic nanostructure that has shown four orders of magnitude fluorescence enhancements—to enable testing from a drop of blood using a cellphone-based fluorescence imaging device in a hospital or home setting. In this project, all components of the plasmonically enhanced assay will be optimized, a smart phone- compatible multiplex chip will be integrated into a passive, capillarity-driven microfluidics device and its figures- of-merit evaluated using archival patient samples. Finally, we will conduct a clinical study where we will quantify the levels of NTpro-BNP, cTnT, and CRP simultaneously from a single drop of blood in approximately 90 patients, employing our point-of-care assay platform, which will be compared to results from commercial immunoassay analyzers at Duke University Hospital. The proposed project is expected to lead to an inexpensive point-of-care cardiac panel, which will have major impact on the diagnosis and treatment of cardiovascular disorders, as detection of multiple biomarkers provides deeper insight into the underlying pathophysiological stages, and self-testing by patients at home enables closer monitoring of treatment response. Moreover, this assay platform can target any analyte for which antibody reagents are available, making it broadly applicable to most, if not all, immunoassay targets in clinical medicine.

PROJECT NARRATIVE The proposed research will develop an inkjet-printed, plasmonically enhanced, single-step, multiplex point-of- care test for the quantification of cardiovascular biomarkers to enable testing from a drop of blood using a cellphone based fluorescence imaging device in a hospital or home setting. This is expected to have a major impact on the diagnosis and treatment of cardiovascular disorders, as detection of multiple biomarkers provides deeper insight into the underlying pathophysiological stages, and self-testing by patients at home enables closer monitoring of treatment response. Moreover, this assay platform can target any analyte for which antibody reagents are available, making it broadly applicable to most, if not all, immunoassay targets in clinical medicine.