The objective of the present study was using short sequence repeats markers (SSRs) in early sexing of the Nile tilapia (Oreochromis niloticus, L.). Molecular markers identification in fishes includes SSRs markers were very useful for investigating of sex determination system, sex-related genes and sex chromosomes. Fin-clip tissue of examined fish was used for DNA extraction and for the amplification of SSRs and sex determination; twenty one primer pairs were tested. The results showed that, from the tested 21 primers of SSRs markers, there were four primers had significant differences between males and females. It was observed that females were homozygous and males may be heterozygous or homozygous for the alternative bands. The females were homozygous and males were heterozygous for two SSRs markers and the females were homozygous and males were homozygous for the alternative alleles for other two markers. In conclusion, the results of the present study and the previous ones confirm the possibility of using short sequence repeats markers (SSRs) in sex determination of fish species include Nile tilapia (Oreochromis niloticus, L.). SSRs markers could be used in the early sexing detection of the Nile tilapia breeding programs and for production supper-male YY without needing for progeny test.