The purpose of this work was to study the effects of electron beam irradiation on development and reproduction and evaluate the DNA damage in Plutella xylostella. Adults and pupae of P. xylostella were irradiated with 30, 50 and 100 Gy electron beam. Hatchability and fecundity of adults declined as increased irradiation doses. When pupae were irradiated with 100 Gy, fecundity of emerged adults significantly decreased and no eggs hatched. However, the adults longevity and emergence of pupae did not change. Assessment of DNA damage in cells obtained from adults and pupae of P. xylostella was carried out using single-cell electrophoresis (comet assay). Electron beam-radiated adults and pupae showed that tail length and percentage of DNA damage at all the doses were significantly larger than the control batch. Our results suggest that electron beam induces sterility through the DNA damage and this technique could contribute to analytical identification of an effective disinfestation and quarantine treatment.