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Electrospinning technology is considered to be one of the efficient, convenient, and low-cost technologies for preparing nanofibers, which can be applied in various industries, including filtration, catalysis, and energy. Here, aiming at the performance requirements of the nanometer fiber filter membrane for filtering PM2.5, the preparation of the nanofiber filter membrane was realized based on multi-needle electrospinning equipment. Meanwhile, by adding silver nitrate to the spinning solution, a polyvinylidene fluoride antibacterial nanofiber filter layer with high filtration efficiency and low resistance was successfully prepared on the traditional air conditioning filter meshes. We found that four key factors affecting the filtration performance include ambient humidity, substrate meshes, voltage, and production rate. Among them, voltage and production rate are the key factors affecting the filtration performance. According to the development trend of multifunctional nanofiber membranes, the preparation of air conditioning filters with nanofibers as the main filter material was realized, producing air conditioning filter membranes with antibacterial and deodorizing functions. This article provides a reliable experimental basis and empirical reference for the preparation of nanofiber membranes based on multi-needle electrospinning technology.