A new method for production of nylon nanofibers with antibacterial properties containing silver nanoparticles (nylon nanofibers/Ag NPs) is introduced via in situ synthesis of nano-silver by reduction of silver nitrate in the polymer solution prior to electrospinning. The properties of the electrospinning solutions and the structures of the electrospun fibers were studied using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDAX), UV-vis spectrophotometer and reflection spectrophotometer. Further, the antibacterial properties of the nanofibers were investigated against Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive) bacteria. Interestingly, an antibacterial properties has been found on nylon 6 nanofibers while the nylon nanofibers/Ag NPs showed excellent antibacterial activities against both tested bacteria. The produced nylon nanofibers/Ag NPs can be a good candidate for biomedical applications, water and air filtration.