Background Meningitis is an acute inflammation of the pia, arachnoids, and the fluid in the subarachnoid space of the brain. Because there is no diagnostic biomarker, it is difficult to diagnose. Objective: The aim of the current study was to evaluate the validity of using cerebrospinal fluid (CSF) Vitamin-D Binding Protein (VDBP) as a new potential marker for diagnosing meningitis. Patients and methods: A cross-section study was conducted on 48 patients with manifestations suggesting an acute meningitis, 28 patients with an acute meningitis who were divided into bacterial group containing 10 patients and viral group containing 18 patients confirmed by laboratory investigations and 20 patients who were clinically suspected as an acute meningitis, but excluded by laboratory investigations, the study was conducted within the period from May 2022 to December 2022. CSF and blood samples were obtained in pairs. CSF and serum VDBP were measured in the 3 groups. CSF VDBP concentrations were compared versus serum VDBP concentrations according to disease (viral meningitis vs. bacterial meningitis vs non-meningitis). Receiver operating characteristic (ROC) analysis for diagnosing meningitis using CSF VDBP concentration was performed. Results: There was a statistical significant difference as regard the CSF VDBP (P<0.01) was found between viral, bacterial and control groups (2.49±.65, 2.43±.55 and 1.74±.25μg/mL, respectively). There was a statistical significant difference (P<0.05) in serum VDBP between the viral, bacterial and control groups (214.5±36.5, 197.4±54.8 and 174.3±40.4 μg/mL, respectively). ROC curve analysis showed that the optimum cut-off level of CSF VDBP for diagnosing meningitis was 1.94 μg/mL with a sensitivity of 82.1% and specificity of 85%. AUC of CSF VDBP was 0.865 (95% CI: 0.761–0.969). Conclusion: The cerebrospinal fluid (CSF) Vitamin-D Binding Protein (VDBP)level demonstrated an excellent diagnostic performance. It could serve as a potential marker for diagnosing acute meningitis.