Chest pain is one of the most common reasons for presentation to the Emergency Department and the ability to rapidly and correctly diagnose the minority of patients who have a myocardial infarction is of critical importance. We assessed the diagnostic performance of a multimarker strategy using heart-type fatty acid binding protein (H-FABP) in combination with a contemporary sensitive troponin (cTn) assay. We measured H-FABP (Randox) and a sensitive cTn (TnI-Ultra, Siemens) at baseline in 343 patients with chest pain enrolled in the prospective BWH-TIMI ED chest pain study. Final presenting diagnosis was adjudicated using all diagnostic data, including the local cTnI results, but reviewers were blinded to H-FABP and the sensitive cTn assays. The diagnostic accuracy of H-FABP and local cTn together (AUC 0.962) was superior to local cTn alone (AUC 0.910, p = 0.0009) with an especially marked improvement in early presenters (AUC 0.983 vs. 0.840, p = 0.0098). In contrast, when combined with the sensitive cTn assay, there was no significant difference in the AUC with H-FABP as compared with the sensitive cTn alone, either in the overall cohort (AUC 0.963 vs. 0.956, p = 0.23) or in early presenters (AUC 0.999 for both). In early presenters, the addition of H-FABP resulted in a NPV of 100% when combined with either the local or sensitive cTn assay. In our study, the addition of H-FABP significantly enhanced the sensitivity and accuracy of diagnosis as compared to a prior-generation troponin assay alone, especially in patients who presented early. H-FABP but did improve overall diagnostic accuracy when added to a current-generation sensitive troponin assay; however, their combination offered the best NPV in early presenters. Further studies are needed to determine the utility a very rapid "rule out" of MI with a single blood draw of troponin and H-FABP at presentation.