ObjectivesMyofascial trigger points (MTrPs) are potential contributors to shoulder pain and can lead to local ischemia and hypoxia, thus causing pain. Color Doppler ultrasound (US) has been used to examine the vascular environment around MTrPs, but has not been used to examine blood flow impairments in patients with shoulder pain and MTrPs. The reliability of color Doppler US for measuring infraspinatus muscle blood flow also has not been established. This study aimed to investigate differences in blood flow between individuals with and without shoulder pain and the reliability of Doppler US for measuring infraspinatus muscle blood flow.MethodsForty participants were enrolled, 20 with shoulder pain and MTrPs and 20 without. Color Doppler US examination was performed twice on each participant to measure peak systolic velocity (PSV), end‐diastolic velocity (EDV), resistive index (RI), and pulsatile index (PI) of the infraspinatus muscle.ResultsThe symptomatic participants had significantly higher PSV (ie, impaired blood flow) than the asymptomatic participants. There were no significant between‐group differences in EDV, RI, and PI. The results also demonstrated good‐to‐excellent intra‐rater reliability for color Doppler US measurements of PSV, EDV, RI, and PI for both groups.ConclusionThis study demonstrated differences in blood flow near MTrPs in the infraspinatus muscle between individuals with and without shoulder pain. It also established good‐to‐excellent reliability of color Doppler US in measuring infraspinatus muscle blood flow. These findings suggest color Doppler US is a useful tool to identify vascular impairments for shoulder pain associated with MTrPs.