The bactericidal properties of copper oxide nanoparticles have growing interest due to potential application in the medical area. The present research investigates the influence of sodium dodecyl sulfate (SDS) and poly(vinylpyrrolidone) (PVP) on the production of copper oxide nanoparticles prepared from copper sulfate (CuSO4) and sodium borohydride (NaBH4) solutions. Different analytical techniques were used to determine the crystal nature, mean size diameter, and surface morphology of the copper oxide nanoparticles. The X-ray diffraction (XRD) patterns showed formation of nanoparticles of cuprite (Cu2O) and tenorite (CuO) when PVP and SDS were added at the beginning of the reaction. In fact, when the Cu/PVP ratio was 1.62, Cu2O nanoparticles were obtained. In addition, nanoparticles of CuO were synthesized when the Cu/PVP ratios were 0.54 and 0.81. On the other hand, a mixture of copper oxides (CuO and Cu2O) and cuprite (Cu2O) was obtained when PVP (Cu/PVP = 0.81 and 1.62) and SDS (Cu/SDS = 0.90) were added 30 min after the beginning of the reaction. Transmission electron microscopy (TEM) images show agglomerated nanoparticles with a size distribution ranging from 2 to 60 nm, while individual particles have sizes between 4.1 ± 1.9 and 41.6 ± 12.8 nm. The Kirby-Bauer method for the determination of antibacterial activity shows that small CuO (4.1 ± 1.9 nm) and Cu2O (8.5 ± 5.3 nm) nanoparticles inhibit the growth of Escherichia coli, Staphylococcus aureus MRSA, S. aureus and Pseudomonas aeruginosa bacteria. The antibacterial test of cotton fabric impregnated with nanoparticles shows positive results. The determination of the optimal ratio of copper oxide nanoparticles per cm2 of fabric that are able to exhibit a good antibacterial activity is ongoing.