At present, tick control methods mainly include chemical control, genetic control, immune control, and biological control; however, chemical control still plays a dominant role in practical production. With ticks responding to selective pressure by various acaricides, the indiscriminate use of pesticides has led to the development of resistance. In present study, laboratory-based larval packet tests (LPTs) were performed to assess chemical acaricides efficacy against Rhipicephalus microplus and Haemaphysalis longicornis ticks collected from fifteen Chinese localities. Overall, Ticks from different regions have developed varying levels (ranged from susceptible to level IV) of resistance to different insecticides. A standard sensitive strain of H. longicornis from China has been established. Comparative alignment of VGSCs gene (domain-II) showed 29 single nucleotide polymorphisms (SNPs) including 5 nonsynonymous (NS) SNPs. Comparative alignment of gamma-aminobutyric acid (GABA)-gated chloride channel gene showed 21 SNPs.