Background: Nearly 3 million newborns suffer from sepsis globally each year and sepsis is considered one of the leading causes of morbidity and mortality in neonates, despite recent advances in health care units. The main burden of the problem falls on the developing world. Objective: The current study aimed to evaluate the prevalence of Neonatal sepsis among newborns unit and antimicrobials is used in government and private hospitals in Sana’a, Yemen. Methods: The current study aimed to evaluate the prevalence of Neonatal sepsis among newborns unit and antimicrobials is used in government and private hospitals in Sana’a, Yemen. A cross-sectional study was conducted on (200) children suspected of having neonatal sepsis from both hospitals (100government, 100 private) in the period from (1January, 2024 until 30April, 2024). Medical information on the newborns was collected using a structured questionnaire. The sample has been examined of members to confirm the growth of the pathogen, its type, and the degree sensitivity and resistance to the antibiotics that were used during treatment. Data were collected and analysed using (SPSS.V25) program and tested using Chi square to verify the extent of relationships between variables within the study at the significance level (P<0.005). Results: The results showed that the prevalence of neonatal sepsis was 107 (53%), the EOS was higher 74(69.2%) than LOS 33(30.8%), and there is a significant effect (p=0.047(of the prevalence of the neonatal sepsis in government hospitals (56.1%) higher than in private hospitals. Most of the pathogens were of the GP bacteria (50.5%), the most common bacteria were (Staph.spp), (49.5%) followed by (Pseudomonas.aeraginosa, E. coli and Klebsiella.spp) bacteria, respectively (15.9%, 10.3%, 9.3%). Most patients with EOS has the highest infection with GN bacteria (55.4), while LOS has the highest infection with GP with statistically significant differences (p<0.05), the prevalence of GP bacteria in hospitals Government (65.96%) higher than private hospitals, with statistically significant differences (p<0.05). The highest bacterial resistance was to (Amoxicillin) at a rate of (100%), followed by (Cefuroxime, piperacillin and Ceftriaxone) with rates respectively (98.7%, 92.8%, 90%), while bacterial resistance was moderate to the both drugs (Cefazidime and Cefepime) with rates respectively (63,.6%,66%), and the least bacterial resistance to the drug (Meropenem) was 1 (1%). There was a statistically significant effect (p<0.05) of bacterial resistance (GN–GP) to both drugs (Gentamycin, Piperacillin, amoxicillin, Augmentin and Cefepime). Conclusion: The prevalence of Neonatal sepsis in government hospitals was higher than in private hospitals, which indicates the need to improve the health situation in government health facilities, and researchers recommend conducting more studies and using broader samples of the groups most exposed to the disease and studying Antibiotic-resistant bacterial pathogen.