The purpose of this study is to investigate microbial contamination in water purifiers from two universities (A and B) in Chungcheong region and to evaluate about the harmfulness of the isolated bacteria to the human. The degree of microbiological contamination of six water purifiers at university A was investigated three times from July 2018 to September 2019, and nine water purifiers at university B were investigated in 2023. The isolated bacteria were biochemically identified using an API kit and Vitek-2 system, and then the bacteria were identified to the species level using MALDI-TOF MS. In addition, the possibility of human infection of the isolated bacteria was evaluated through a literature search. In July 2018 and September 2019, the number of bacteria isolated inside the faucet was below the acceptable standard for hot water, but exceed for cold water in all water purifiers. In January and September 2019, bacteria exceeding the acceptable standards were isolated nine times from the cold water of six water purifies (a total of 12 water purifiers). Bacteria identified by MALDI-TOF MS included anaerobic bacteria (Clostridium novyi, Clostridium themopalmarium etc.), Gram-positive bacilli (Microbacterium testaceum, Arthrobacter woluwensis etc.), and Gramnegative bacilli (Acinetobacter nosocomialis, Comamonas kerstersii etc.), which are difficult identify by biochemical methods. In conclusion, bacteria exceeding the acceptable standard were isolated from the cold water of most of the water purifiers. Most of the isolated bacteria were low-pathogenic bacteria from natural environment, but opportunistic bacteria that can cause infection in humans were also isolated from some water purifiers.