Fourteen sets of microsatellites primers were tested on randomly selected individuals of great sturgeon, H. huso. Five sets were developed for lake sturgeon (Afu-19, 34, 39, 54, and 68) and seven sets were developed for Adriatic sturgeon (An-0, 1, 16, 20, 40, 76, and 77) and f nally two sets were developed for Chinese sturgeon (As-73, and 74). Except two sets of Adriatic sturgeon primers, An-76, and An-16, the rest of primers reproducibly amplif ed the beluga sturgeon DNA. Five pairs of primers out of 14 have been selected and conf rmed to be used for separating the populations. Those primers are; Afu-19, Afu-39, Afu-54 from lake sturgeon; As-73 from Chinese stur geon, and An-77 from Adriatic sturgeon. The samples from the east region of the southern Caspian Sea did not amplify at Afu-19, Afu-39, Afu-54, and An-77 loci. However, samples from middle and west regions of the Caspian Sea did amplify at Afu-19, Afu-39, Afu-54, An-77 loci in different manners of producing the bands. Samples from all three regions did amplify at As-77 locus. However, the bands at the samples from the east region are distributed dif ferently from the bands at th e samples from the middle and west regions. Statistical analysis revealed that the number of alleles per locus ranged from 6 to 1one locus had at least six alleles. Population differentiation combination test (Fisher's method) based on both genic differentiation and on genotypic differentiation (G-based) for the two sets of samples demonstrated that the probabilities are highly signif cant. It means possibly two great sturgeon populations exist in the southern part of the Caspian Sea