Reports on the influence of the stress on giant magnetoimpedance (GMI) effect of CoFeSiB films are rarely seen. Therefore, the influence of stress on the GMI effect of CoFeSiB thin-film was studied in this investigation. Firstly, to select suitable substrates for applying stress, CoFeSiB films were fabricated on different flexible substrates with magnetron sputtering, and the hysteresis loops of the films were measured. The results show that the magnetic properties of the films on polyester (PET) substrates and silicate flexible glass are similar and better than polyimide (PI). So they are candidate materials for flexible substrates. And then, the GMI curves of the films under different stresses were measured. The results show that applying longitudinal tensile stress would reduce the longitudinal anisotropy and weaken the GMI effect of the CoFeSiB film. The research on the influence of stress on GMI thin-film would be helpful to fabricate flexible GMI sensors of stable performance.