Abstract: Objective To establish a rapid,sensitive and specific diagnostic test for detecting Neisseria gonorrhoea.Methods The major outer membrane proteins(P_Ⅰ)in different gonococcal serogroups were obtained by isolation of outer membrane complex with CTB-ethanol precipitation,the outer membrane proteins were extracted with Z 3,14 and EDTA,and purified with DEAE-Sepharose CL-6B to obtain P_Ⅰ.Hybridoma cell lines producing McAbs against P_Ⅰ were established with lymphocyte hybridoma techniques.Results The molecular weight of P_ⅠA and P_ⅠB were determined with SDS-PAGE as 35.2k Da and 36.7k Da,respectively.Five hybridoma cell lines producing McAbs continuouslly and stably against P_ⅠA and P_ⅠB were obtained,in-cluding two hybridoma cell lines producing McAbs against P_Ⅰ A and three hybridoma cell lines producing McAbs against P_Ⅰ B.The titers of McAbs in the supernatants in the cultures and in abdominal ascites of BALB/c were from 1:64to1:256 and from 1:4096 to 1:16384,respectively;and the specificity of the McAbs against P_ⅠA and P_Ⅰ B was so high that they easily reacted with N.gonorrhoeae but did not with other antigens such as N.meningitidis etc.Conclusion The purified P_Ⅰ and the McAbs obtained in the study provide a basis to establish a rapid,sensitive and specific diagnostic test for detecting N.gonorrhoea.

Key words: Neisseria gonorrhoeae, Oter membrane proteins, bacterial, Antibodies, monoclonal