A multiple-antigen detection assay for tuberculosis diagnosis based on broadly reactive polyclonal antibodies

Dai, Zhenhua and Liu, Zhiqiang and Xiu, Bingshui and Yang, Xiqin and Zhao, Ping and Zhang, Xuhui and Duan, Cuimi and Que, Haiping and Zhang, Heqiu and Feng, Xiaoyan (2017) A multiple-antigen detection assay for tuberculosis diagnosis based on broadly reactive polyclonal antibodies. Iranian Journal of Basic Medical Sciences, 20 (4). pp. 360-367.

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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.

Item Type: Article
Subjects: QW Microbiology and Immunology
Divisions: Journals > Iranian J Basic Medical Sciences
Depositing User: ijbms ijbms
Date Deposited: 22 Sep 2017 22:25
Last Modified: 22 Sep 2017 22:25
URI: http://eprints.mums.ac.ir/id/eprint/2502

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