Insecticide resistance amongst agricultural pests and disease vectors are an increasing threat to human health, and the production of fruits and vegetables with the overuse of pesticides is causing serious ecological damage and environmental contamination. The breed of strains with resistance to insects has become an important laboratory approach in studies on resistance mechanisms and insect-pest control, and has gained growing attention from scientists worldwide. Research on breeding strains with resistance to flies, which are one of the widespread agricultural and public health pests, can also provide reliable data and the theoretical basis to understand the resistance mechanism of harmful flies and for future pest management. The present review performed a systematic summary on the following fields. First, we review the resistance mechanisms of flies and other insects, exploring physiological, biochemical, behavioral, genetic variation, and symbiotic resistance. Second, we review four cultivation method frequently applied for flies, such as topical application, dipping, residual film and spraying. Third, we summarize the identification method and influencing factors of resistant strains, such as virulence determination, resistance multiplier calculation, gene expression, and genetic variation. Finally, we compare the recent identification and breeding resistant varieties method . In conclusion , this review aims to provide significant application value and research insights for studies of resistance mechanisms, resistance monitoring and integrated pest management.