Introduction: Acute Myocardial Infarction (AMI) is a serious life threatening condition having a high mortality and morbidity rate. Hence, early detection and appropriate treatment is essential. Clinically Troponin I and Creatinine Kinase Myocardial Band (CK-MB) are currently used to detect AMI. But its rise in blood concentrations is seen only after 4-6 hours after the onset of AMI. Clinical research shows that Heart-type Fatty Acid Binding Protein (H-FABP), a novel biomarker, is beneficial in the early detection of AMI. Aim: To determine the diagnostic accuracy of H-FABP and compare with the existing blood biochemical markers, such as Troponin I and CK-MB, in the early detection of AMI in south Indian population. Materials and Methods: This cross-sectional study was conducted in Sree Balaji Medical College and Hospital, south Chennai, Tamil Nadu, India, from March 2017 to April 2019. A total of 50 participants, aged between 40-75 years, and diagnosed with ST elevation myocardial infarction presenting within 24 hours to the emergency room were included. Serum was collected during the time of admission (0 hour), between 4-6 hours from the time of admission and between 12-24 hours from the time of admission for estimation of cardiac specific biomarkers such as cardiac troponin I, H-FABP, CK-MB. The sensitivity, specificity and accuracy of the cardiac-specific biomarkers were calculated using Receiving Operating Characteristics (ROC) curve analysis. Results: There were 38 males and 12 females, with a mean age of 62.1±4.7 years. The sensitivity of H-FABP at the time of admission (0 hour) was 88%, 78% between 4-6 hours of admission, and 42% between 12-24 hours of admission. At 0 hour, the sensitivity of H-FABP (88%) was high compared to cardiac troponin I (43%) and CK-MB (40%). The accuracy of H-FABP and cTnI was equal (80% each) at the time of admission. At 4-6 hours after admission, cardiac troponin I showed higher sensitivity, specificity and accuracy compared to H-FABP. ROC analysis showed the Area Under the Curve (AUC) for H-FABP was higher (0.81) compared to cardiac troponin I and CK-MB. Conclusion: H-FABP was found to be more sensitive compared to cardiac troponin I or CK-MB in AMI patients. Hence, early diagnosis of AMI is made possible with the H-FABP measurements.