Abstract: To develope a rapid and accurate test for detecting serotype O foot-and-mouth disease virus (FMDV)based on the principle of gold immunochromatographic assay(GICA),guinea pig and rabbit polyclonal antibody against FMDV serotype O were purified by DEAE-sephrose.The guinea pig anti (FMDV) serotype O-IgG was conjugated with colloidal gold ,which was called gold probes,and was dispensed on the glass fiber. The rabbit anti (FMDV) serotype O-IgG and the goat anti-guinea pig IgG polyclonal antibody were separately sprayed on the nitrocellulose membrane as the test line (T line) and the control line (C line). The rapid gold immunochromatographic strip was assembled by the different accessory in regular sequence. If there was serotype O FMDV in sample, gold probes on the glass fiber and rabbit anti (FMDV) serotype O-IgG formed a sandwich complex with serotype O FMDV.Within the test line, the sandwich complex was immobilized, furthermore, concentrated and appeared a distinct red color on the test line.In the clinic test, a total of 53 field samples were comparatively detected with both GICA and reverse indirect hemagglutination assay (RIHA), and the positive rate were 95.45% and 90.91%, respectively.The evaluation test proved GICA established in this study is simple, rapid, sensitive and specific.It is suitable for basic veterinary medical laboratories.