Abstract Background Presenteeism refers to being present at work but experiencing reduced productivity due to health problems, and has been known to be related to sleep loss. Workers commonly sleep longer on days off than on workdays, and presenteeism may be reduced with extended sleep on days off. Aims This study aimed to determine the association between sleep duration both on workdays and days off and presenteeism. Methods The participants were 1967 workers who engaged in work for 5 days and rested for 2 days weekly. Sleep duration was classified into less than 6 hours (short; S), 6–8 hours (medium; M), and 9 hours or longer (long; L), for workdays and days off, respectively. Presenteeism was assessed using the World Health Organization Health and Work Performance Questionnaire. Results On both workdays and days off, compared to medium sleep duration, short sleep duration was significantly associated with increased odds of presenteeism. The odds of presenteeism were significantly increased for S-S (odds ratio [OR] 2.17, 95% confidence interval [CI]1.40–3.37), S-M (OR 1.59, 95% CI 1.14–2.22), S-L (OR 2.71, 95% CI 1.05–7.00), and M-S (OR 6.82, 95% CI 2.71–17.17) combined sleep duration for workdays and days off, respectively, compared to an M-M (reference). Conclusions Sleep loss on workdays cannot be compensated for with longer sleep on days off. This study suggests that sufficient sleep duration on both workdays and days off is important for reducing presenteeism.