The aim was to analyze the relationship between periods with uninterrupted neck muscle activity for ≥4 min and neck and shoulder pain. The trapezius muscle activity was recorded bilaterally on 40 young workers and students during a full shift. Neck and shoulder pain, mechanical work load and decision control were reported at the same time as the muscle activity recording and 6 months later. A dose-response relationship was found between uninterrupted muscle activity and neck and shoulder pain, with a ten-fold higher risk for the group with more than half, compared to less than a third, of the shift with uninterrupted muscle activity. Self-reported mechanical work load showed a small but protective effect related to pain. Gender and decision control did not emerge as important risk factors in this model. In conclusion, this study indicates that work or other exposures that contains long periods with uninterrupted neck muscle activity of 4 min duration or longer should be minimized to reduce risk of neck and shoulder pain.