Background: Early detection and start of antibiotic therapy neonatal sepsis (N.S) dramatically improves outcomes, so it is important to perform fast, reliable and specific early laboratory biomarkers. Aim: This study aimed to detect the prevelance, the risk factors, hematology profile, microbial profile of neonatal sepsis patients and also investigate the value of PCT and CRP, in comparison to presepsin in establishing the early diagnosis of neonatal sepsis. Methods: A cross sectional study was performed from March to September 2019 in Al Quwayiyah General hospital involving 120 neonates who were classified into 3 groups. The patients groups Khater and Al-Hosiny; JAMB, 20(1): 80-90, 2020; Article no.JAMB.54569 81 were: Proved N.S, suspected N.S and control healthy neonates, classified depending on Tollner score. Haematology profile and blood culture for each neonate were done. CRP, PCT and presepsin values were analyzed, compared, and their effectiveness as diagnostic markers was determined. Sensitivity, specificity, positive, and negative predictive values of the markers were calculated. Results: The prevelance of neonatal sepsis was 20.8%. 75 neonates were males and 45 neonates were females. 74 neonates were preterm, while 46 were full term. Gestational age in weeks was 31.1±5.9w for neonates with proved sepsis, 32.4±6.7w for neonates with suspected sepsis and 36.4±4.4w for control group. The mean birth weight was 1740±105.3 g for neonates with proved sepsis, 32.4±6.7 g for neonates with suspected sepsis, 2.650±205.2 g for control group. 36 babies suffered from respiratory distress syndrome, 10 had jaundice, 8 had cough, 28 had fever and 8 complained of other symptoms. Blood cultures were positive for all patients of proved sepsis. The identified bacteria included Gram positive bacteria 22(55%) which were Coagulase negative staph. 13(32.5%) followed by Staphylococcus aureus 4(10%) while Gram negative bacteria 15(37.5%) which were E. coli 5(12.5%) followed by Klebsiella peumoniae and also fungal infection (Candida species) detected in 3(7.5%) cases. There was significant difference between the mean and standared deviation of CRP, PCT and presepsin levels in proved and suspected N.S. groups when compared with healthy controls (P< 0.05). CRP sensitivity and specificity (72%, 61% respectively) which were less useful in diagnosis of neonatal sepsis compared to presepsin which has the highest sensitivity and specificity (95%, 81% respectively) followed by procalcitonin with sensitivity and specificity (90%, 69% respectively). Conclusion: The prevalence of neonatal sepsis among all admitted neonates in Al-Quwayiyah general hospital was 20.8%. Our results also detected higher sensitivity, specificity and positive and negative predictive values for presepsin more than and PCT CRP in the diagnosis of NS. Original Research Article