It is difficult to identify the sex of Ostrich ( Struthio camelus) with morphological method before reaching sexual maturity,and this situation challenged the early feeding of Ostrich. In this study,genome DNA was extracted from feathers by using nondestructive sampling method and a combination of primer from one of EE0. 6 and the other of CHD. Sex gene sequences were amplified and compared among three pairs of sex-known birds and 9 sex-unknown birds. The results showed that male birds displayed no distinct amplification band, whereas one band was amplified for female ones. Thus the method could be used for sexing Ostrich before sexual maturity.