The invention provides a method for preparing copper oxide antibiosis fibers. The method comprises the following steps: 1, carrying out fusion mixing on copper oxide powder, a silicon-containing surfactant and polyester slices to prepare functional master batches, wherein the diameter of the copper oxide powder is smaller than 0.8[mu]m, and the functional master batches comprise, by mass, 30-40% of the copper oxide powder and 4-6% of the silicon-containing surfactant; and 2, mixing the functional master batches with the polyester slices, adding the obtained mixture to a screw extruder, and spinning to obtain the copper-containing antibiosis fibers. An organosilicon quaternary ammonium salt and a cationic silicon-containing surfactant are used in fabrics in the invention, so the fabrics have good softness, resilience and smoothness, and an obvious antistatic effect, and the fibers obtained by commonly using the organosilicon quaternary ammonium salt and copper oxide have better spinnability and antibiosis than fibers obtained by individually using copper oxide.