This study aimed to provide data on high-quality cardiopulmonary resuscitation (CPR) by determining the fatigue effects on rescuers wearing level D personal protective equipment (PPE) while performing CPR. The study’s methodology entailed the comparative performance of two-person CPR; once while the participants wore level D PPE and once while they did not, over the 30 min course in seven cycles. The fatigues of 24 students from the department of emergency medical technology were measured pre-experiment and during CPR, after each cycle was measured. In this study, there were no differences in the blood lactate concentration, heart rate, oxygen saturation and subjective fatigue participants with or without level D PPE over the course of repeated CPR cycles. However, participants wearing level D PPE typically displayed the ‘somewhat hard’ and ‘hard’ stages of subjective fatigue a cycle faster than those not wearing level D PPE. Specifically, the proportion of such participants who reached more than the ‘somewhat hard’ stage of fatigue in the second cycle was higher than that among those not wearing level D PPE (p < .05). Consequently, wearing level D PPE was found to increase the rate of subjective fatigue from the early course of performing CPR. Therefore, rescuers will be able to provide higher quality CPR if measures are conducted to reduce fatigue.