Electrospinning is favored as a continuous and efficient method for preparing nanofibers. The purpose of this study is to combine polyamide (PA) with Zinc oxide (ZnO) through electrospinning and uniform precipitation method to obtain ZnO/PA composite nanofibers with excellent conductivity, antibacterial property and cell compatibility. The morphology and structure of ZnO/PA composite nanofibers were analyzed and the effects of process parameters on the morphology of ZnO crystals were investigated. Conductivity of ZnO/PA composite nanofibers in wetting condition was measured using a four-probe resistance tester. The antibacterial activity of ZnO/PA composite nanofibers was tested by inhibition zone method, and MG-63 human osteosarcoma cells were used to detect the cytotoxicological effects of the composite nanofibers. The results show that the petal-like ZnO crystals are densely and evenly distributed on the surface of nanofibers, and the thickness of the ZnO crystal is 38.77 ± 2.42 nm. Conductivity of ZnO/PA composite nanofibers in wet state reaches (1.064 ± 0.0189) × 10-3 S∙mm−1, significantly higher than that of PA nanofibers (about (1.486 ± 0.017) × 10-4 S∙mm−1). The ZnO/PA composite nanofibers have good antibacterial activity against Escherichia coli and Staphylococcus aureus, and exhibit good cytocompatibility on MG-63 cells.