Background: Neonatal sepsis (NS) is an important cause of morbidity and mortality among newborns. Its diagnosis depends mainly on blood culture that takes at least 48 hours to give results. Therefore, searching for biomarkers for early diagnosis is of value. We aimed to assess presepsin, soluble triggering receptor expressed on myeloid cells (sTREM-1), and neutrophil CD64 (nCD64) as early diagnostic biomarkers in NS, and to compare them individually and in combination. Methods: This hospital-based case-control study has been conducted on 60 full-term neonates recruited from the neonatal intensive care unit, Al-Zahraa Hospital, Al-Azhar University, Cairo, Egypt. Thirty infants with sepsis were compared to 30 postnatal age- and sex-matched healthy controls. Studied neonates were evaluated using clinical and laboratory indicators for sepsis. nCD64 was measured by flow cytometry and, serum presepsin and sTREM-1 were measured by ELISA. Results: Presepsin, sTREM-1, and nCD64 levels were significantly elevated in septic neonates vs control group (P<0.05). The sensitivities of presepsin, sTREM, and nCD64 were 100%, 96.7%, and 86.7%, respectively. Presepsin had the best diagnostic performance in early diagnosis of NS followed by sTREM-1 and nCD64. Conclusion: Presepsin and sTREM-1 are promising biomarkers in screening for NS in comparison with nCD64. However, nCD64 is better used in combination with other biomarkers as CRP.Keywords: diagnosis; nCD64; neonatal sepsis; presepsin; sTREM-1; screening.