Recently a biosafety level-3(BL-3) mobile laboratory has been set up for the virus scanning and vaccine development because of the COVID-19 pandemic. The study on air flow inlet and outlet location and its flow direction with ventilation in the mobile laboratory needs to prevent spread of COVID-19 virus because the COVID-19 virus is primarily transmitted to people through respiratory droplets and aerosol coming out as their coughing. This study is conducted on the air flow pattern optimization in BL-3 mobile laboratory with various design specifications of position of air supply & exhaust port and particle source. Air flow patterns of ceiling supply-exhaust and ceiling supply-bottom side exhaust with particle source were determined to compare the impact of the infection prevention. CFD simulation was used to analyze for two air flow patterns and particle source position. Numerical results showed that air flow pattern of air conditioning system with ceiling supply-exhaust in a row is more effective than that of ceiling supply-bottom side exhaust air flow pattern in terms of infection prevention in biosafety mobile laboratory.