Background: Delayed neurological sequels (DNS) have been described after carbon monoxide (CO) poisoning. There is a need to find a new prognostic marker to guide the use of hyperbaric oxygen (HBO) therapy. Aim: To evaluate serum S-100β level in patients presenting with acute CO poisoning as an indicator of poisoning severity and predictor of DNS occurrence and HBO need in those patients. Methods: This prospective cohort study included patients with acute CO poisoning. On admission, carboxyhemoglobin (COHb) and S-100β levels were measured. Patients were followed up for 6 months for signs of DNS. Results: Out of 50 patients, 6 only developed DNS. The mean of S-100β levels was significantly higher in patients with severe poisoning and those with DNS. Receiver operating characteristic curve analysis revealed that S-100β had an area under the curve 0. 871; at a cutoff value ≥ 0.67 µg/L, its sensitivity and specificity were 100% and 77.3%, respectively. The sensitivity of S-100β was significantly higher than that of COHb, while its specificity and overall accuracy were significantly higher than those of HBO criteria. Conclusion: Serum S-100β level on admission could be a marker of poisoning severity and a predictor of CO-induced DNS development that guides the use of HBO therapy.