Abstract: In order to establish a rapid immunological detection method of naphthalene and other polycyclic aromatic hydrocarbons (PAHs), which originated from leakage of petroleum transportation in the marine, highly efficient and stable hybridoma cell line secreting anti-naphthalene McAb is urgently needed.In this study, 2-naphthyl butyric acid -KLH was used as immune antigen to immunize BALb/c mice in order to detect serum titer in mice coupled with indirect competitive ELISA. The technique of cell hybridoma screening was used to screen hybridoma cell line resistant to anti-naphthalene monoclonal antibody and determined the immunoglobulin subtype of mice with capture ELISA.Results showed that the mice serum titer can reach the maximum value of 1:80000.The five cell lines which can secrete anti-naphthalene monoclonal antibody after selection by HAT, and subclones whose cell supernatant titer can reach a maximum value of 1:15625.All of the immunoglobulin subtypes of anti\aphthalene monoclonal antibody were IgG1, Kappa light chain.As a result, the successfully constructed hybridoma cell lines can stably secrete anti-naphthalene monoclonal antibody, whose antibody subtypes agree with the immunological detection characteristics.Our method provides as a stable and efficient source of monoclonal antibody for establishing the detecting method of ELISA as well as a reference for developing the colloidal gold immunochromatographic method for ELISA detection of naphthalene (GICA) strip.