Objective To understand pathogenic microorganism contamination status of medical waste sharps containers with different use time, explore the reasonable duration of service time of sharps containers, provide reference for the management of medical waste. Methods Twelve 2L sharps containers on treatment trolleys in a tertiary first-class infectious disease specialty hospital were randomly selected, viral loads of hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV) as well as bacterial colonies on inner and outer surfaces of sharps containers at different time points were detected, three unused sharps containers were taken as control at the same time. Results Sixty eluent specimens of outer surface and contents of sharps containers in trial group and control group were collected respectively at four time points (48 h, 72 h, 5 d, 7 d), no HIV and HCV were detected, and no HBV was detected in specimens of outer surface of sharps containers, HBV was detected in the eluent of contents in one sharps container 72 hours after the use, concentration of HBV was 2.20E+01 IU/mL. Changes in bacteria in the eluent of used sharps containers:100% of the eluent of contents in sharps containers grew bacteria on the 5th day after use, bacterial load of the eluent of contents in sharps containers on the 7th day after use was incalculable. Bacterial load on the outer surface of sharps containers ranged from 1 to 9 CFU/cm². No significant changes were observed in the inner and outer surfaces of all sharps containers, and no discomfort odor emerged. Conclusion With the storage time prolonged to 7 days, bacterial colonies on the outer surface of sharps containers didn't increase significantly, HIV, HBV and HCV were not detected. It is suggested that service time of sharps containers with small production of contents should not be set compulsorily at 48 hours (even if the contents in sharps container is less than 3/4 of storage capacity after 48 hours of use).