This study examined the relationship between the peak running velocity (PRV) obtained during a horizontal, incremental treadmill test and distance running performance in a group of highly-trained male (N = 14) and female (N = 9) distance runners. Performance (5 km run time) was assessed with a self-paced time trial under laboratory conditions in an attempt to minimize extraneous variables which could affect performance (i.e. environmental conditions, terrain, etc.); relationships with recent best 5 km race time were also determined. PRV was highly related to 5 km performance whether determined from the time trial (r2 = 0.94, p < 0.001) or recent race (r2 = 0.89, p < 0.001). A new finding was that PRV was similarly related to performance in both the male (r2 = 0.83, p < 0.001) and female (r2 = 0.80, p < 0.001) athletes. Peak running velocity is thus highly predictive of distance running performance in highly-trained endurance runners. This finding has important practical implications, as PRV can be measured without extensive metabolic equipment and/or invasive techniques.