Background:
Developing countries often lack biosafety standard operating procedures. A need to study biosafety practices among staff of state and private laboratories has been identified. This studyaimedto assess the knowledge and practices of laboratory standard precautions (LSP) among laboratory staff in in human and animal laboratoriesin the governorate of Tunis.

Methods:
A cross-sectional descriptive study were conducted using a standardized questionnaire administered between December 2021 and January 2022. Data were collected among laboratory staff who agreed to participate in this study. Answers were judged according to the score granted to each item and used during a face-to-face interview.

Results:
Among the152 staff interviewed, 70% were from the public sector. The majority of staff was vaccinated against hepatitis B (84.5%). The overall level of knowledge was satisfactory in 51% of cases. In the area ofproper use of personal protective equipment, 95% wore the mask and 80% the gloves. On a practical level, 66% of the staff questioned, regardless of their categories, had an adequate overall perception of biosafety.

Conclusion:
Although this study revealed several limitations such as the reluctance and unavailability of laboratory staff during the study period, significant gaps affecting both knowledge and practice among laboratory staff in Tunis region have been identified. Continuous staff training based on behavior and on application of universal and specific hygiene measures as well as periodic evaluations of these laboratories is recommended.