With the rapid growth of fresh-cut produce market, the South Korean fresh-cut industry is facing the challenge of ensuring food safety. As the estimation of the microbial numbers in fresh-cut produce processing lines (tools, and equipment) using the conventional microbiological techniques takes days, so there is a need for faster and easier monitoring methods. This study was conducted to investigate the use of ATP bioluminescence assay to measure the degree of microbial contamination from three actual fresh-cut processing lines. The samples collected from frech-cut vegetables, and fresh-cut fruits processing plants were tested for the estimation of the bacterial number, using the ATP bioluminescence and microbiological methods. The result of former was transferred to log RLU/100 cm2, and that of the latter was transferred to log CFU/100 cm2. A positive linear correlation between the ATP bioluminescence assay value and aerobic-plate count was found for fresh-cut processing lines, with a correlation coefficient of 0.8772 (n=50). The results of this study indicate that ATP bioluminescence assay can be used to monitor microbial contamination in fresh-cut produce processing plants, and can help improve the hygiene therein.