A large number of team sports require athletes to repeatedly produce maximal or near maximal sprint efforts of short duration interspersed with longer recovery periods of submaximal intensity. This type of team sport activity can be characterized as prolonged, high-intensity, intermittent running (PHIIR). The primary purpose of the present study was to determine the physiological factors that best relate to a generic PHIIR simulation that reflects team sport running activity. The second purpose of this study was to determine the relationship between common performance tests and the generic PHIIR simulation. Following a familiarization session, 16 moderately trained (JOURNAL/jscr/04.02/00124278-200702000-00025/ENTITY_OV0312/v/2017-07-20T235329Z/r/image-pngO2max = 40.0 ± 4.3 ml·kg−1·min−1) women team sport athletes performed various physiological, anthropometrical, and performance tests and a 30-minute PHIIR sport simulation on a nonmotorized tread-mill. The mean heart rate and blood lactate concentration during the PHIIR sport simulation were 164 ± 6 b·min−1 and 8.2 ± 3.3 mmol·L−1, respectively. Linear regression demonstrated significant relationships between the PHIIR sport simulation distance and running velocity attained at a blood lactate concentration of 4 mmol·L−1 (LT) (r = 0.77, p < 0.05), 5 × 6-second repeated cycle sprint work (r = 0.56, p < 0.05), 30-second Win-gate test (r = 0.61, p < 0.05), peak aerobic running velocity (Vmax)(r = 0.69, p < 0.05), and Yo-Yo Intermittent Recovery Test (Yo-Yo IR1) distance (r = 0.50, p < 0.05), respectively. These results indicate that an increased LT is associated with improved PHIIR performance and that PHIIR performance may be monitored by determining Yo-Yo IR1 performance, 5 × 6-second repeated sprint cycle test work, 30-second Wingate test performance, Vmax, or LT. We suggest that training programs should focus on improving both LT and Vmax for increasing PHIIR performance in moderately trained women. Future studies should examine optimal training methods for improving these capacities in team sport athletes.