The aim of this study is to simulate the transportation/temporary storage, frozen storage, and reuse scenarios of infectious specimens in public health biological sample banks. By adding Escherichia coli to rabbit serum and drinking water to simulate infectious human serum samples and environmental water samples, the effects of different transportation temperatures, freezing temperatures, and freeze-thaw cycles on bacterial activity in samples, especially in deep low-temperature environments, were evaluated. The results showed that under transportation conditions of 4 ℃ for infectious serum and drinking water, the number of Escherichia coli in the samples gradually decreased over time. Under dry ice transportation conditions, the number of Escherichia coli in the drinking water samples decreased to an extremely low level after 12 h. Under the conditions of －20 ℃， －80 ℃， and liquid nitrogen, there was no statistical difference between the results of Escherichia coli count after 1 to 3 times freeze-thaws of serum and the reference value. This study suggests that key data affecting sample quality at each stage should be recorded throughout the entire process, and pre-analysis information such as transportation temperature, time limit, and total number of freeze-thaw cycles should be recorded in the biological sample library information system. Although they are not biological sample operation information, the accuracy of the results can be directly affected.