BACKGROUND: Workers in the microbiology laboratories are continuously exposed to the risk of laboratory-associated infections. Tuberculosis (TB) is a frequent laboratory-acquired infection owing to production of cough-generated aerosols with ease and high infectivity of Mycobacterium tuberculosis. This study aims to investigate the current situation of biosafety in Korean TB laboratories. METHODS: We conducted a nationwide survey of laboratories in hospitals conducting TB tests using questionnaires about their facility and management standards. RESULTS: We analyzed data from 52 hospitals nationwide that have a capacity of 100–2,000 beds, of which only two laboratories conduct high risk drug-susceptibility testing on cultured isolates among other test items, whereas six laboratories perform only direct sputum-smear microscopy. The remaining laboratories performed moderate-risk activities/tests, like sample processing for culture. In the majority of these laboratories, there are laboratory medicine specialists who are fully in charge of health checkup programs for laboratorians. The facility and management standards vary widely according to the size of the hospital and risk of TB tests. CONCLUSIONS: Our survey results about the current situation of TB laboratories could be useful as baseline data for preparing biosafety guidelines for all TB laboratories in Korea.