Brown dog tick, Rhipicephalus sanguineus sensu lato, an important ectoparasite transmitting several pathogens, is the most common tick species infesting dogs. Control of ticks being central to the control of fatal tick-borne diseases, this study attempted to assess the susceptibility/resistance of brown dog ticks to synthetic pyrethroids, the commonly used acaricides against ticks. Larval packet assay revealed 60% of isolates tested to be resistant and tolerant to deltamethrin as per the resistance factor that ranged from 1 to 53.7. Sequence analysis of the PCR amplified product of domain II S4-5 linker of sodium channel gene in R. sanguineus revealed novel polymorphisms, viz., C190A, G215T and T270C. In domain III S6 region of the gene, a T2134C mutation was observed. Genotyping with allele-specific PCR targeting domain II S4-5 linker region using single larvae revealed that most R. sanguineus larvae in the study population were homozygous resistant (RR) genotypes, followed by heterozygous (RS) and homozygous susceptible (SS) genotypes. A higher proportion of RS genotypes was also observed in domain III S6 region. This first report of genotyping of Indian R. sanguineus to analyse synthetic pyrethroid resistance highlights the need to devise alternate control strategies to reduce the brown dog tick population.