Nanostructured and conducting polymer polyaniline (PANI) is used in numerous applications in electrotherapy, electro-magnetic materials for monitoring health, antimicrobial clothing, data transfer in smart textiles, biosensors and for defense technology. An important criterion for all the above mentioned utilities is, producing polymeric conductive fibers. In the present study we prepared conducting PANI nanofibres combined with mupirocin, a topical antimicrobial agent, through a self-assembly process. The prepared polymer was then tested for the antibacterial properties against various Gram positive and Gram negative bacteria such as Streptococcus pyogenes, Staphylococcus epidermidis, Staphylococcus aureus and Escherichia coli. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) were used to identify the chemical structure of the PANI nanofibres. The antibacterial properties were assessed by measuring the zones of inhibition. It was evident from these results that antimicrobial activity increased with increasing PANI and PANI combined with mupirocin (PANI-mupirocin) concentrations. It was also found that PANI-mupirocin has enhanced antimicrobial activity compared to PANI alone. This information might be useful to evaluate the potential use of nanostructured polyaniline in fabrics incorporated with antibacterial agents as a prophylactic use against bacterial skin infections in the near future.