Methicillin-resistant Staphylococcus aureus (MRSA) is an important nosocomial pathogen that shows resistance to many antibiotics and is usually associated with serious infections. Having the ability for biofilm formation increases resistance to antibiotics. Sanguisorba officinalis L. is a perennial plant that is distributed in the northern districts of China and has been used as a traditional Chinese medicine. In this study, the effect of S. officinalis on MRSA strain SA3 isolated from a dairy cow with mastitis was evaluated by testing the growth and biofilm formation ability of MRSA cultured with or without ethanol extracts of S. officinalis. The results showed that the ethanol extract of S. officinalis strongly inhibited the biofilm formation of MRSA. With a confocal laser scanning microscope system, we observed that the biofilm structure of the test group with the addition of S. officinalis appeared looser and had less biomass compared with the control group without S. officinalis. Furthermore, we found that the transcript levels of the icaADBC operon remarkably decreased upon addition of the ethanol extract of S. officinalis, indicating that S. officinalis inhibits biofilm formation of MRSA in an ica-dependent manner.