Here, we have investigated in vitro antimicrobial efficacy of a quaternized cationic polymer, poly[bis(2-chloroethyl)ether-alt-1,3-bis[3-(dimethylamino)propyl]urea] (polyquaternium-2), against gram-positive as well as gram-negative bacteria along with several multi-drug-resistant bacterial strains. The antimicrobial efficacy of this polymer was first tested against some clinical pathogens followed by microorganisms isolated from acne lesions. Interestingly, polyquaternium-2 exhibited significant antimicrobial activity against methicillin-resistant Staphylococcus aureus, for which very limited drugs are available. Most importantly, the polymer displayed low haemolytic activity and non-toxic behaviour against mammalian cells. The results showed the promising potential of the projected polymer to be utilized as an antibacterial agent for various biomedical applications.