Cardiovascular diseases are fast growing problems since last century and vast studies has proven helpful to combat the problems more frequently but yet no tool could be discover to control the cardiovascular problems at early stages. Myeloperoxidase traditionally known as an inflammatory enzyme released from leukocytes during inflammation of cells. This book therefore, provide a new success to overcome the problem related to diagnosis of cardiovascular problems, more cost effective method as compared to traditional methods use currently. Present study is concerned with the development of enzyme linked immunosorbant assay (ELISA) for myeloperoxidase (MPO). Assay was validated by recovery and dilution experiment and show high % recovery and enough sensitivity as all requires. Second part of the present study was clinical application of developed assay. Blood serum MPO levels were measured from healthy individuals both male and female and results were compared with serum MPO levels of patients suffering from Acute Coronary Syndrome. The results were highly significant p> 0.001, showing that this test could be used for research and diagnosis of cardiovascular diseases.