Previous studies have shown improved external chest compression (ECC) quality and delayed rescuer fatigue when the dominant hand (DH) was in contact with the sternum. However, many rescuers prefer placing the non-dominant hand (NH) in contact with the sternum during ECC. We aimed to investigate the effects of up-down hand position switch on the quality of ECC and the fatigue of rescuers during cardiopulmonary resuscitation (CPR). After completion of a review of the standard adult basic life support (BLS) course, every candidate performed 10 cycles of single adult CPR twice on an adult manikin with either a constant hand position (CH) or a switched hand position (SH) in random order at 7-day intervals. The rescuers' general characteristics, hand positions, physiological signs, fatigue appearance and ECC qualities were recorded. Our results showed no significant differences in chest compression quality for the DH position rescuers between the CH and SH sessions (p>0.05, resp.). And also no significant differences were found for Borg score (p = 0.437) or cycle number (p = 0.127) of fatigue appearance after chest compressions between the two sessions. However, for NH position rescuers, the appearance of fatigue was delayed (p = 0.046), with a lower Borg score in the SH session (12.67 ± 2.03) compared to the CH session (13.33 ± 1.95) (p = 0.011). Moreover, the compression depth was significantly greater in the SH session (39.3 ± 7.2 mm) compared to the CH session (36.3 ± 8.1 mm) (p = 0.015). Our data suggest that the up-down hand position switch during CPR may delay the fatigue of non-dominant hand position rescuers and improve the quality of chest compressions.