Background: Sepsis has a poor prognosis for critically ill patients, even with intensive management. Early diagnosis of sepsis and detection of patients with worsening prognosis are important for immediate intervention to improve the clinical outcome. Objective: To investigate serum presepsin (PS) and procalcitonin (PCT) as early diagnostic and prognostic biomarkers for sepsis in critically ill patients. Methods: 60 critically ill patients with sepsis were subdivided into three groups of sepsis, severe sepsis and septic shock according to Acute Physiology and Chronic Health Evaluation II (APACHEII) and quick Sequential Organ Failure Assessment (qSOFA) scores. Patients were compared with 20 age and sex matched controls. Serum PS and PCT were measured by enzyme linked immunosorbent assay (ELISA). Results: Serum PS and PCT levels were significantly increased in septic patients than controls, and their increase was positively correlated with progression of sepsis severity till reached the highest levels in septic shock. Receiver operating characteristic (ROC) curve for predicting sepsis revealed that PS has the highest area under curve (AUC) (0.967) with 97.5% sensitivity, 85% specificity and cut-off of >635.5 pg/mL, followed by PCT that has AUC (0.946), 97.5% sensitivity, 95% specificity and cut-off of >319.7 pg/mL. C-reactive protein (CRP) showed the lowest AUC (0.902) with 75% sensitivity, 100% specificity and cut-off of >7 mg/L. ROC curve for predicting septic shock showed that PS has the highest AUC (0.969) with 90% sensitivity, 97.5% specificity and cut-off of >5500.6 pg/mL, followed by CRP that has AUC (0.945), 90% sensitivity, 87.5% specificity and cut-off of >63 mg/L. PCT showed the lowest AUC (0.889) with 90% sensitivity, 97.5% specificity and cut-off of >822.1 pg/mL. Conclusions: Serum PS and PCT were promising biomarkers for early diagnosis and prognosis of sepsis in critically ill patients, but PS was superior to PCT.