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This study aims to evaluate the biosafety awareness of laboratory staff working on pathogens detection in seven Centers for Disease Control and Prevention and support these staff\'s biosafety management and training. A total of 208 laboratory staff from seven provincial CDCs were chosen by stratified random sampling to undergo a self-administered questionnaire survey. The collected data were analyzed using SPSS 22.0. The overall average biosafety awareness of the CDC laboratory staff involved in pathogen detection was 82.5 points. The average biosafety awareness score was the highest in health monitoring (92.63 points) and the lowest in risk assessment and control (41.6 points). Among the seven provincial CDCs, the Guizhou Provincial Center for Disease Control and Prevention laboratory staff showed the lowest average biosafety awareness score (74.3 points). The team who worked for 5-14 years were more likely to score above the passing score (≥88 points; corrected OR 0.400, 95%CI: 0.168-0.951) than the staff with less than five years of work experience. In addition, the mid-level and junior-level staff, as well as the lower position ones were more likely to score below the passing score (<88 points; junior level and lower: corrected OR 3.473, 95%CI: 1.028-11.737; mid-level: corrected OR 2.797, 95%CI: 1.027-7.618) compared with the senior-level staff. Among the surveyed team, 66.3% identified a lack of specific funds related to work, 61.5% identified a lack of designated personnel for the laboratory biosafety management. The biosafety awareness in CDC laboratory staff involved in pathogen detection is low, especially regarding risk assessment and control. The biosafety awareness in Guizhou and Guangxi Provincial CDC laboratory staff is also low. Laboratory funding, job title, and years of experience in a laboratory influence biosafety awareness in CDC laboratory staff. The biosafety knowledge, education, and training of CDC laboratory staff involved in pathogen detection need to improve by paying attention to the content and coverage of biosafety training, exploring new training modalities, and increasing funding for activities related to biosafety in CDC.“,”This study aims to evaluate the biosafety awareness of laboratory staff working on pathogens detection in seven Centers for Disease Control and Prevention and support these staff\'s biosafety management and training. A total of 208 laboratory staff from seven provincial CDCs were chosen by stratified random sampling to undergo a self-administered questionnaire survey. The collected data were analyzed using SPSS 22.0. The overall average biosafety awareness of the CDC laboratory staff involved in pathogen detection was 82.5 points. The average biosafety awareness score was the highest in health monitoring (92.63 points) and the lowest in risk assessment and control (41.6 points). Among the seven provincial CDCs, the Guizhou Provincial Center for Disease Control and Prevention laboratory staff showed the lowest average biosafety awareness score (74.3 points). The team who worked for 5-14 years were more likely to score above the passing score (≥88 points; corrected OR 0.400, 95%CI: 0.168-0.951) than the staff with less than five years of work experience. In addition, the mid-level and junior-level staff, as well as the lower position ones were more likely to score below the passing score (<88 points; junior level and lower: corrected OR 3.473, 95%CI: 1.028-11.737; mid-level: corrected OR 2.797, 95%CI: 1.027-7.618) compared with the senior-level staff. Among the surveyed team, 66.3% identified a lack of specific funds related to work, 61.5% identified a lack of designated personnel for the laboratory biosafety management. The biosafety awareness in CDC laboratory staff involved in pathogen detection is low, especially regarding risk assessment and control. The biosafety awareness in Guizhou and Guangxi Provincial CDC laboratory staff is also low. Laboratory funding, job title, and years of experience in a laboratory influence biosafety awareness in CDC laboratory staff. The biosafety knowledge, education, and training of CDC laboratory staff involved in pathogen detection need to improve by paying attention to the content and coverage of biosafety training, exploring new training modalities, and increasing funding for activities related to biosafety in CDC.