Hydrogen peroxide gas sterilization is a widely used low temperature sterilization technique, yet still with limitations in material compatibility and penetration, resulting in a high failure rate of biological monitoring. Through review of literatures, this paper comprehensively and deeply explores the causes and mechanisms of failures in biological monitoring by hydrogen peroxide gas sterilization (including true positive, false positive, and false fluorescence negative reaction), helps professionals analyze and judge the causes of failures in biological monitoring, and provides ideas and references for reducing failure rate.