关键词: 广州管圆线虫, 可溶性抗原, 单克隆抗体, 循环抗原, 夹心ELISA法

Abstract: 　Objective To develop and identify monoclonal antibodies (McAbs) against adult worm of Angiostrongylus cantonensis and observe its applicability. Methods BALB/c mice were immunized with soluble antigen of adult worms of A.cantonensis. The spleen cells of immunized mice were fused with myeloma cell, and the hybridoma secreting high titer of McAbs with high specificity was screened. By using the McAbs, serum of angiostrongyliasis patient and sera of the rats infected with A.cantonensis were detected by Western blotting and double antibody sandwich ELISA respectively. Results Three McAbs were established (2A2,3F1,4H2), which all showed no cross reaction with antigens of Schistosoma japonicum, Paragonimus westermani, Cysticercus cellulosae and Trichinella spiralis. Western blotting analysis demonstrated that the three McAbs recognized a Mr 15 000 soluble antigen of adult worm of A.cantonensis and recognized the Mr 24 000 and Mr 15 000 circulating antigens from the serum of angiostrongyliasis patient. The double antibody sandwich ELISA detection showed a positive rate of 76.5%. Conclusion Three hybridoma cell lines against adult worm of A.cantonensis have been established which secret high titer of McAbs with high specificity and seem promising in detecting the circulating antigen of the angiostrongyliasis patient.

Key words: Angiostrongylus cantonensis, Soluble antigen, Monoclonal antibody, Circulating antigen, Sandwich ELISA