Abstract: As BSL-2 laboratories are established to have the highest level of biosafety in most disease control agencies at basic level, the uncertainties of species of pathogenic microbes in tested samples in routine investigation of disease and the demand for practicality, feasibility and safety, the present paper provides a detailed elaboration on the position, layouts, enclosure, ventilation, environmental parameters, security equipments and personal protection of BSL-2 laboratories in disease control agencies based on classification, summarization, and interpretation of applicable laws/regulations on biosafety laboratory construction and personal experience to provide the reference for the construction or reconstruction of BSL-2 laboratory