Genetic diversity was studied among 40 different zone samples in the Indian short-nosed fruit bat, Cynopterus sphinx. Samples were collected from 127 locations in different districts of southern Tamil Nadu, India. Polymorphism at molecular level was studied by random amplified polymorphic DNA marker technique. Polymerase chain reaction (PCR) with 30 arbitrary decamer oligonucleotide primers was applied to the 40 zone samples. Out of 30 random primers screened, 22 gave reproducible polymorphic bands. PCR product of bat genome revealed a total of 234 bands, out of which 171 were found to be polymorphic. The dendrogram displaying the relative genetic similarities between the zones ranged from 0.42 to 0.92. Principal coordinates analysis revealed that the first two components accounted for 51.4% of the total variation. The results show that considerable genetic diversity was observed in this species from different zonal populations, possibly due to complete dispersal of juveniles of both sexes from their natal groups and gene flow between the zones. The results obtained from this study suggest not only a predictive framework for future studies, but also the use of genetic data in the management and meaningful conservation of this species