Low-level biosafety laboratories (BSL-1 and BSL-2) are widely established in medical, research, and educational institutions and represent the most common settings for pathogenic microorganism-related experimental activities. Although these laboratories mainly handle pathogens with low to moderate hazards, they still pose biosafety risks due to frequent experimental operations, high personnel mobility, and substantial variations in management practices. In recent years, laboratory-associated infection incidents have indicated that low-level biosafety laboratories constitute a weak link in biosafety management. Based on a systematic review of relevant domestic and international studies and policy documents, this paper summarizes the risk characteristics, major risk types, and risk assessment methods of low-level biosafety laboratories. Particular attention is given to biological exposure risks, risks associated with personnel operations and behaviors, and management-related risks. In addition, the application progress of qualitative, quantitative, and semi-quantitative risk assessment methods in low-level laboratories is compared and analyzed. Furthermore, existing challenges in current risk assessment practices, such as the lack of unified indicator systems and insufficient application of assessment results, are discussed, and future directions for risk assessment in low-level biosafety laboratories are proposed. This review provides references for improving risk assessment systems and enhancing biosafety management in low-level biosafety laboratories.