›› 2006, Vol. 24 ›› Issue (5): 2-332.

• 论著 • Previous Articles     Next Articles

Preparation of Specific Antibodies Against Recombinant Em18Antigen and Detection of the Circulating Antigen

JIANG Li1;LI Xiong2;LI Hao1;NIU Xin-ling2;FENG Zheng1   

  1. 1 National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention,WHO Collaborating Center for Malaria,Schistosomiasis and Filariasis,Shanghai 200025,China;2 Xinjiang Center for Disease Con-trol and Prevention,Urumqi 830002,China;* Shanghai Municipal Center for Disease Control and Prevention, Shanghai,200336,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2006-10-30 Published:2006-10-30
  • Contact: JIANG Li

Abstract: Objective To prepare specific antibodies against the recombinant Em18 antigen (ReEm18), estab-lish a sandwich ELISA and detect the Em18 circulating antigen in patients, sera. Methods Rabbits and BALB/c mice were immunized with the ReEm18 antigen, which was purified by affinity method for preparation of the specific poly-clonal and monoclonal antibodies. A sandwich ELISA was established by the specific antibodies. Results By immu-nizing with the ReEm18 antigen, high antibody level was reached with a serum dilution of 1 ∶ 204 800 and above in the immunized rabbits. After double selection by ELISA using the ReEm18 antigen and block ELISA using both AE-positive and negative control sera, 14 positive cell clones were obtained with an inhibition rate of more than 50%. Those mono- and poly-clonal antibodies were matched freely in sandwich ELISA tests for detecting the ReEm18 antigen. A combina-tion of monoclonal antibody No.9 and polyclonal antibody showed the best result. The sensitivity to detect ReEm18 anti-gen was at 3 ng/ml. Six of 11 AE sera were positive when tested with the sandwich ELISA system. Conclusion Highly specific polyclonal and monoclonal antibodies have been prepared, and a sensitive sandwich ELISA established. Preliminary result is suggested that a detectable level of Em18 circulating antigen is present in AE patients, sera.

Key words: Alveolar echinococcosis, Em18 recombinant antigen, Antibody, Circulating antigen, Sandwich ELISA