The aim of this longitudinal study was to assess the effect of rotating shift work on perceived sleep quality and sleep duration of nurses at Queen Elizabeth Central Hospital, Blantyre, Malawi. Twenty four female nurses were recruited at random from among personnel engaged in rotating shift work. The nurses worked a three-phase schedule: five day shifts (7.00 -17.00) followed by three night shifts (17.00 -7.00) and five days off. Controls were 22 female nurses who did not perform night duties. Sleep quality and duration was assessed using standardized and validated questionnaires on sleep duration and subjective sleep quality (SSQ). One-way analysis of variance revealed a significant effect of shift phase on total sleep duration (F = 36.8, d.f. = 8, P < 0.000) and perceived sleep quality (F = 8.81, d.f. = 3, P < 0.000). Night shift work was associated with reduction of sleep quality and duration. The after effects of night shifts persisted during days of the recovery period indicating accumulation of fatigue.