Potato virus Y (PVY) is distributed around the world and causes an important economic loss. We report here, the development of a recombinant antibody in the form of single chain variable fragment (scFv) against PVY by using phage display technology. A library of single chain variable fragments was constructed by using the antibody variable region (V) genes of B cells derived from the spleens of PVY immunized Balb/CJ mice. The selection of scFv with ability to bind PVY was performed by biopanning and tested against PVY by phage-enzyme linked immunosorbent assay (ELISA). ELISA tests showed that scFvs produced by five clones from randomly selected 44 clones reacted specifically with PVY. These five clones didn't show cross reaction with tobacco mosaic virus(TMV). These results suggest that the application of phage display technology can be used for developing immunodiagnostic kits for detection of PVY and trangenic plant against PVY.