Objective This study aimed to evaluate soluble cluster of differentiation 14 subtype (sCD14-ST), also named presepsin, as an early marker for the diagnosis of culture-proven early-onset sepsis (EOS) in neonates and to assess its relation to disease severity and mortality. Study Design Out of 60 neonates with risk factors of EOS, 31 neonates were diagnosed as having culture-proven EOS. They were compared with 20 nonseptic controls. We obtained blood samples on day 1 of life for sCD14-ST measurement and sepsis screening. Blood samples were repeated on day 3 in EOS neonates. Results sCD14-ST was significantly higher in EOS neonates than controls (p < 0.001). Neonates who later developed septic shock had significantly higher day 1 sCD14-ST level than those who did not (p < 0.001). Furthermore, neonates who died had significantly higher day 1 sCD14-ST than survivors (p < 0.001). On day 3, there was a significant decline in sCD14-ST levels than initial levels among survivors. There was a significant positive correlation between day 1 sCD14-ST level and Tollner's sepsis severity score. Conclusion sCD14-ST could be used as a powerful diagnostic and prognostic marker of EOS. Its quantitative measurement at birth could be a good predictor of sepsis severity and mortality.