Thiocyanates, especially bis-alkylthiocyanates are highly effective in killing a number of bacterial strains and are reported to be potent biocides at ppm concentrations. In order to examine whether a covalently bound and immobilized thiocyanate group on a biomaterial surface is still effective as a bactericide, plasticized poly(vinyl chloride) (PVC) was thiocyanated using sodium thiocyanate in the presence of a phase transfer catalyst in aqueous media leading to the nucleophilic substitution of chlorine by thiocyanate on the PVC surface. Thiocyanation imparted hydrophilicity to the surface in comparison with bare PVC. Control and thiocyanated PVC surfaces were exposed to two strains of bacteria commonly implicated in device-associated infections, such as Staphylococcus aureus and Staphylococcus epidermidis. Bacterial adhesion and colonization was quantitated by counting the viable organisms on the adhered surface as well as by optical and scanning electron microscopy. Significantly reduced retention of S. epidermidis and S. aureus was seen on the thiocyanated PVC surface. Immobilized thiocyanate was non-cytotoxic in a preliminary cell culture assay. The study thus showed that even though an immobilized thiocyanate moiety on the polymer surface was not as effective as a bactericide unlike soluble thiocyanates, it prevented the retention and colonization of the bacteria to a considerable extent.