Diffusion-weighted imaging (DWI) with single-shot echo-planar imaging (ssEPI) is a valuable tool for detecting acute brain lesions but does suffer from image distortions. Multishot echo-planar imaging (msEPI) is a technique for reducing such distortions. This study aimed to compare the image quality and diagnostic efficacy of ssEPI- and msEPI-DWI at 5.0 T for brain disease detection. This study retrospectively reviewed images of 107 consecutive patients with suspected brain diseases who underwent ssEPI- and msEPI-DWI at 5.0 T at the First Affiliated Hospital of University of Science and Technology of China from August 2023 to September 2023. Two radiologists independently graded image quality and measured the image distortion. Signal-to-noise ratio, contrast-to-noise ratio, and apparent diffusion coefficient (ADC) were calculated and compared between ssEPI- and msEPI-DWI. Image quality scores were compared using the Wilcoxon test and other continuous variables by the paired t-test. The diagnostic accuracy of ADC values in distinguishing lesions from normal-appearing tissues was measured with the area under the curve (AUC). Image quality evaluation and distortion analysis revealed that msEPI-DWI significantly outperformed ssEPI-DWI (two-sided P<0.001). No significant difference was observed in signal-to-noise ratio, contrast-to-noise ratio, or ADC values between msEPI- and ssEPI-DWI (two-sided P≥0.601). The ADC values of msEPI- and ssEPI-DWI showed strong correlations for both lesions (r=0.97) and contralateral normal tissues (r=0.91) (two-sided P<0.001). Compared to those of the contralateral white matter, ADC values of low-grade gliomas (LGGs) were significantly higher [ssEPI-DWI: 1,119.9±273.1 vs. 805.1±73.9; msEPI-DWI: 1,196.2±355.6 vs. 757.3±98.0 (unit: ×10-6 mm2/s)], while the ADC values of acute cerebral infarction (ACI) lesions were significantly lower [ssEPI-DWI: 603.9±273.2 vs. 888.9±212.0; msEPI-DWI: 538.0±281.2 vs. 905.0±188.9 (unit: ×10-6 mm2/s)] (two-sided P≤0.003). The AUCs for detecting LGGs were excellent for both ssEPI-DWI [AUC =0.934; 95% confidence interval (CI): 0.84-1.00] and msEPI-DWI (AUC =0.944; 95% CI: 0.86-1.00) (two-sided P<0.001; two-sided DeLong test: P=0.833). As compared to ssEPI-DWI, msEPI-DWI, when performed at 5.0 T, demonstrated superior image quality and less anatomical distortion in a wide spectrum of brain diseases and showed promising diagnostic performance for LGGs and ACI. In the future, msEPI-DWI at 5.0 T could become clinically routine in the diagnosis and grading of brain disorders.