Background: Despite numerous brain magnetic resonance imaging (MRI) utilization in the emergency department (ED), certainly, imaging alone is not enough, and it is necessary to have a correct interpretation by a physician who has sufficient skills in this regard. Objectives: Here, we decided to investigate the accuracy of interpreting brain MRI of suspected acute ischemic stroke (AIS) patients conducted by emergency medicine physicians (EMPs) in comparison with a radiologist and a neurologist. Methods: This diagnostic accuracy study was conducted from April to November 2019 in Tehran, Iran. All attending EMPs of one major educational, medical center, a radiologist, and a neurologist also participated. A set of brain MRI stereotypes related to patients suspected of having a AIS was randomly selected. By reviewing the brain MRI interpretation of EMPs, once in comparison with the radiologist and once in comparison with the neurologist, misinterpretations (presence or absence of findings compatible with the diagnosis of AIS) were extracted. Results: Brain MRI stereotypes of 287 suspected AIS patients were interpreted of these patients, 160 cases (55.7%) were male. The mean age of the study patients was 65.0 ± 14.1 (range of 18 to 98) years. The value of the agreement for diagnosis between EMPs and neurologists was 0.684 (95% CI: 0.580 to 0.787). Considering the neurologist as the gold standard, the accuracy of AIS diagnosis by the EMPs was 0.85% (95% CI: 79.3 to 89.6). The agreement value for diagnosis between EMPs and radiologist was 0.673 (95% CI: 0.553 to 0.794). Considering the radiologist as the gold standard, the accuracy of AIS diagnosis by the EMPs was 86.3% (95% CI: 79.8 to 91.3). The agreement value for diagnosis in these two groups was 0.752 (95% CI: 0.627 to 0.877). Conclusions: The findings of the current study revealed that the accuracy of brain MRI interpretation performed by the EMPs, compared with both neurologist and radiologist was proper.