Background: Glioma a common type of the brain tumor, which originates in the glial cells that surround and support neurons in the brain, including astrocytes, oligodendrocytes and ependymalcells. Objectives: This study is aiming to investigate the role of MRS in patients in diagnosis of brain glioma. Patients and methods: The study was designed as a cross-sectional study recruit 30 patients, who were referred to the Radio-diagnosis Department of South Valley University Hospitals for suspected glioma from neurosurgery department during the period from February 2018 to Aug 2021 the study protocol was approved by the local ethics committee of South valley University hospitals (Ref No Code Ms-4–2018, Date of approval 10–02-2018). and it was approved by the local Research Ethics Committee of our hospital. Results: The study included 30 patients with age ranged from six to 72 years old. The lesions within the repetitive tumor group had essentially higher Cho/Cr ratios than those within the radiation damage group (p < 0.0001; SE, 0.18), and these, also, had vitally higher Cho/Cr proportions than the typical white matter of 6 patients (p = 0.0003; SE, 0.09). In differentiating tumor versus non-tumor lesions, the sensitivity and specificity of Cho/NAA ratio greater than 2.0 was 96% and 70%, respectively. For ratios higher than 2.5, the sensitivity and specificity to detect tumors was 97% and 86%, respectively. Conclusion: MRS examination was greatly beneficial in differentiating neoplastic from non-neoplastic lesions and decrease for stereotactic brain biopsy. However, further studies are required to confirm the current evidence.