Acinetobacter calcoaceticus-Acinetobacter baumannii (ACB) complex has emerged as the most common opportunistic nosocomial pathogen. The biofilm producing ability of nosocomial strains play a significant role in health care setting and antimicrobial resistance. A prospective cross-sectional study was performed in the microbiology laboratory of Annapurna Neurological Institute and Allied Science, Kathmandu Nepal, from May to December 2015.