ARGs as emerging environmental pollutants pose a threat to human health, and studies show that secondary effluent from sewage treatment plants is an important source of ARGs in the environment. Ozone pre-oxidation combined with PAC and UF was used to treat the secondary effluent from the wastewater treatment plant, and the factors and mechanisms of O3 dosage, pH, and temperature on the removal of different forms of ARGs in the secondary effluent were investigated. The results show that ARGs could be more effectively removed by PAC adsorption and UF after oxidative inactivation by O3; under the condition of optimal O3 dosage (2.0 mg/L), the combined O3-PAC-UF process could remove different types of ARGs (tetA, tetC, tetG, sulI, sulII) and other pollutants (intⅠ1, 16S rRNA) from the secondary effluent. The removal of different types of ARGs (tetA, tetC, tetG, sulI, sulII) and other pollutants (intⅠ1, 16S rRNA) by the combined process is 102.67~103.92 copies/mL, and the removal of cellular and free ARGs is significantly enhanced; the removal of ARGs is facilitated by the elevated pH and lower temperature; the direct oxidation of O3 molecules dominate the removal of ARGs in the secondary effluent. In conclusion, ozone combined with PAC and UF can effectively remove antibiotic resistance genes from secondary effluent.