Author/Authors :
Dai، Zhenhua نويسنده Department of Bio-diagnosis,Beijing Institute of Basic Medical Sciences,Beijing,China , , Liu، Zhiqiang نويسنده Department of Bio-diagnosis,Beijing Institute of Basic Medical Sciences,Beijing,China , , Xiu، Bingshui نويسنده Department of Bio-diagnosis,Beijing Institute of Basic Medical Sciences,Beijing,China , , Yang، Xiqin نويسنده Department of Bio-diagnosis,Beijing Institute of Basic Medical Sciences,Beijing,China , , Zhao، Ping نويسنده Chaoyang District Centre for Disease Control and Prevention,Beijing,China , , Zhang، Xuhui نويسنده Department of Bio-diagnosis,Beijing Institute of Basic Medical Sciences,Beijing,China , , Duan، Cuimi نويسنده Department of Bio-diagnosis,Beijing Institute of Basic Medical Sciences,Beijing,China , , Que، Haiping نويسنده Department of Bio-diagnosis,Beijing Institute of Basic Medical Sciences,Beijing,China , , Zhang، Heqiu نويسنده Department of Bio-diagnosis,Beijing Institute of Basic Medical Sciences,Beijing,China , , Feng، Xiaoyan نويسنده Department of Bio-diagnosis,Beijing Institute of Basic Medical Sciences,Beijing,China ,
Abstract :
Objective(s): Detection of circulating Mycobacterium tuberculosis (M. tuberculosis) antigens is promising in Tuberculosis (TB) diagnosis. However, not a single antigen marker has been found to be widely expressed in all TB patients. This study is aimed to prepare broadly reactive polyclonal antibodies targeting multiple antigen markers (multi-target antibodies) and evaluate their efficacies in TB diagnosis. Materials and Methods: A fusion gene consisting of 38kD, ESAT6, and CFP10 was constructed and overexpressed. The fusion polyprotein was used as an immunogen to elicit production of multi-target antibodies. Their reactivities were tested. Then, the multi-target antibodies and three corresponding antibodies elicited by each single antigen (mono-target antibodies) were evaluated with sandwich ELISA for detecting M. tuberculosis antigens. Their diagnostic efficacies for TB were also compared. Results: The polyprotein successfully elicited production of multi-target antibodies targeting 38kD, ESAT6, and CFP10 as analyzed by Western blotting. When used as coating antibodies, the multi-target antibodies were more efficient in capturing the three antigens than the corresponding mono-target antibodies. By testing clinical serum, the multi-target antibodies demonstrated significantly higher sensitivity for clinical TB diagnosis than all three mono-target antibodies. Conclusion: The multi-target antibodies allowed detecting multiple antigens simultaneously and significantly enhanced TB detection compared to routine mono-target antibodies. Our study may provide a promising strategy for TB diagnosis.
