【】 Objective: Advances in biotechnology have increasingly highlighted the importance of biosafety within the field of public health. Low-level biosafety laboratories, due to their large numbers and wide range of applications, represent a critical link in biosafety risk management. There is an urgent need for the effective control and prevention of their potential risks.Methods: This study engaged front-line biosafety laboratory personnel from hospitals, Centers for Disease Control and Prevention (CDC), and health authorities as research subjects. A two-round anonymous expert consultation was conducted using the Delphi method. Subsequently, a combined weighting approach integrating the Analytic Hierarchy Process (AHP) and the Entropy Weight Method was applied to establish a comprehensive risk assessment index system for low-level biosafety laboratories that incorporates both subjective and objective elements.Results: The first round of consultation involved 30 experts, and the second round involved 53 experts. Through two rounds of anonymous solicitation and iterative revision, a systematic risk assessment index system for low-level biosafety laboratories was finalized. This system comprises 5 first-level indicators (Personnel Risk Prevention, Equipment Safety Management, Specimen Risk Management, Laboratory System Construction, Laboratory Environment Management), 10 second-level indicators, and 30 third-level indicators. The overall Kendall"s coefficient of concordance was 0.23 (P < 0.001), and the consistency ratios (CR) of all judgment matrices were less than 0.1. These results indicate that the system development process was scientifically rigorous, expert opinions were well-coordinated, and the weight assignments are reliable, making the system suitable for practical risk assessment and management in low-level biosafety laboratories.Innovation and Application Value: Based on the five dimensions of "Personnel, Equipment, Specimen, System, and Environment," this study developed a unified risk assessment index system for low-level biosafety laboratories and quantified the indicator weights. This enriches research on standardized assessment in this field. The system is applicable to various scenarios, including daily risk control in laboratories, supervisory inspections by regulatory bodies, and related academic research.