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Introduction The lifetime risk of developing acute coronary syndrome (ACS) is heavily influenced by modifiable factors like smoking, hypertension, diabetes, and lifestyle, as well as non-modifiable factors such as age, sex, and genetic predisposition. As the primary symptom of ACS, chest pain accounts for the most common reasons for emergency department visits and outpatient cardiac evaluations in the United States, posing a significant diagnostic challenge to clinicians. ACS carries a massive economic burden, with high direct costs from hospitalizations and high indirect costs like lost productivity, impacting healthcare systems, especially in rural areas like West Virginia, with a high prevalence of risk factors associated with cardiovascular disease progression. Circulating biomarkers present a promising approach in the research and clinical practice of various diseases as they are minimally invasive, highly cost-effective, and provide high specificity. So, the objective of the present study was to evaluate the potential of a biomarker panel in differentiating cardiac causes of chest pain in non-ACS patients in West Virginia. Methods The exploratory cross-sectional study was conducted in patients from a hospital in West Virginia, United States. Patients who were referred to the Rapid Access Chest Pain Clinic were enrolled. Patients were selected based on the established inclusion and exclusion criteria. The plasma samples from chest pain patients, in comparison with age-matched controls, were used for the assessment of circulating biomarkers of cardiac dysfunction such as N-terminal pro-B-type natriuretic peptide (NT-proBNP), Myosin-binding protein C (MyBP-C), creatine kinase-myocardial Band (CK-MB), and neopterin. Results Our results showed that there was a significant elevation in these biomarkers in patients with chest pain in comparison with healthy controls. The correlation analysis revealed a significant link between these parameters and NT-proBNP, the well-established and highly specific predictive markers of cardiac injury and cardiac function decline. This multimarker approach that assessed the additive value of the proposed biomarker analysis may provide an earlier assessment of overall patient risk and may aid in identifying patients with a higher risk of an adverse event. Conclusion The findings in the present study demonstrate the potential of the four-biomarker panel in identifying the cardiac causes of chest pain and the future translational applicability of the proposed biomarker panel to determine and compare the prognostic abilities for early ACS detection through detailed longitudinal studies. This may also help in diagnosis, risk stratification, and guidance of treatment, further allowing management of cardiac function decline and improved health outcomes in the high-risk population of West Virginia.