Background: Meningitis represents a critical medical situation that requires appropriate therapy to ensure the patient's survival and recovery. In such cases, differentiation between viral and bacterial meningitis is crucial to determine the most effective course of treatment. S100 calcium binding protein B (S100B) functions as a biomarker for the brain inflammatory process. Neuron-specific enolase (NSE) serves as a biomarker for neuronal stress and neurodegenerative disorders. Objectives: The purpose of this research was to assess S100B and NSE in the serum and cerebrospinal fluid (CSF) of patients with acute meningitis and to assess the diagnostic utility of both markers to identify the cause of meningitis, whether bacterial or viral. Patients and methods: Thiscross-sectional research was conducted on 44 patients, divided into two groups: bacterial meningitis and viral meningitis. S100B and NSE levels in the serum and CSF were determined by the enzyme-linked immunosorbent assay. Biochemical and cytological examination and cultures of CSF were performed. Results: The paraclinical examinations revealed that the bacterial meningitis group exhibited significantly elevated levels of ESR, CRP, WBC count, neutrophils, and urea compared to the viral meningitis group (p < 0.05). Furthermore, as compared to the viral meningitis group, the bacterial meningitis group exhibited significantly higher levels of CSF protein, CSF leukocyte count, and neutrophils (p < 0.05). Bacterial meningitis was associated with significantly higher serum concentrations of S100B than viral meningitis (p=0.047). Conclusion: SerumS100B is a simple, non-invasive biomarker that can be used for the early prediction and diagnosis of acute bacterial meningitis rather than acute viral meningitis.