Abstract: To develop a rapid and effective method for Equine influenza virus (EIV) detection, polyclonal antibodies against EIV A/equine/Xinjiang/07 strain and monoclonal antibody against NP of EIV were generated respectively. Then a double antibody sandwich ELISA (DASELISA) was developed. The specificity of the optimized DASELISA was evaluated using EIV, Equine arteritis virus, Equine herpes virus1, Equine herpes virus4 and Japanese encephalitis virus, resulting in only EIV specimens yielding a strong signal. Compared with hemagglutination test, its sensitivity was as two point five to ten times as the later. And it had crossreactivity with H7N7 subtype. Meanwhile it is suitable for detection of virus from the nasal swabs of experimentally infected equines. The results revealed that the ELISA possessed good specificity and higher sensitivity, indicating a suitable method for rapid detection of EIV.