Background: The correct treatment and management of scabies is expensive and time-consuming, and may have a negative impact on patients and their families. Aim: To investigate the effects of permethrin 5% cream on scabies mites, and explore mite survival times outside the human body. Methods: We performed a nonrandomized controlled study. In total, 20 petri dishes were coated with permethrin 5% cream (treatment group) and 20 plain petri dishes (control group) each had one scabies mite placed in them, and were then observed at baseline and 3, 4, 5, 6, 7, 8 and 12 h from baseline. In the second part of our study, 30 scabies mites from infested patients were investigated in an observational experiment in 30 plain petri dishes at days 0, 3 and 4. Results: Our data showed that 65% of scabies mites survived after 8 h in the treatment group compared with 75% of mites in the control group. After 12 h, 25% of mites in the treatment group and 60% in the control group were still alive. Data from the observational survival study showed that one mite was alive on day 3, but all mites were dead by day 4. Conclusions: This study showed no significant effects of mite survival times with 5% topical permethrin after 8 h, while its efficacy was stronger and significant after 12 h. We recommend the isolation of all mite-infested items for at least 4 days.