Microbial decontamination is a critical concern in various sectors, from healthcare to food processing. Traditional decontamination methods, while effective to a degree, present limitations in terms of environmental impact, efficiency, and potential harm to the target material. This review investigates the emerging realm of non‐thermal plasma (NTP) as a promising alternative for microbial decontamination, emphasizing its mechanisms, reactor designs and applications. The mechanism decomposed into physical, chemical and biological effects of plasma, are elaborated upon to provide a foundational understanding of the intrinsic principles of plasma decontamination. Except for the generation type of NTP, reactors and other parameters by which NTP achieves microbial decontamination, emphasizing the design considerations and parameters that influence its efficacy. Moreover, the latest applications of NTP in decontaminating air, water, and surfaces, supported by the latest research findings in each domain are explored. Additionally, the perspectives on the future research tendencies of NTP decontamination and disinfection are highlighted with potential avenues for exploration and innovation. Through this comprehensive review, the aim is to underscore the potential of NTP, particularly DBD plasma, as a versatile, efficient, and environmentally friendly method for microbial decontamination.