An accurate and rapid assay of cardiac nerve growth factor (NGF) levels in blood can provide physicians with critical information regarding myocardial injury and neural remodeling in cardiac tissues to identify patients at risk of impending heart attack, thereby enabling them to receive appropriate lifesaving treatment more quickly. Currently used assay methods, such as enzyme-linked immunosorbent assay (ELISA), are usually time-consuming (hours to days), expensive and technically complicated. In this paper, we described the development and clinical study of a rapid and sensitive method for detection and quantification of NGF in human blood plasma. This method utilizes a fiber-optic, immuno-biosensing system which performs a fluorophore-mediated sandwich immunoassay on the surface of an optical fiber. Physiological concentrations of NGF could be quantified in both buffer and human blood plasma samples within 5 minutes. The NGF concentrations determined by the fiber-optic sensor were comparable to those by the gold standard, ELISA. Preliminary study of NGF assay in cardiac patient plasma samples showed a great potential of the fiber-optic sensor as a rapid diagnostic and prognostic tool in clinical applications.