Novel monoclonal antibodies to ESAT-6 and CFP-10 antigens for ELISA-based diagnosis of pleural tuberculosis
| Autor: | Lu Hz, Yi-Wei Tang, Feng Tt, Shou Cm, Fan J, Yao Hp, Shen L, Wu Zg, Qian Y, Zhang Yz, Wu Np |
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| Rok vydání: | 2011 |
| Předmět: |
Pulmonary and Respiratory Medicine
Tuberculosis medicine.drug_class Pleural effusion Immunoblotting Enzyme-Linked Immunosorbent Assay Tuberculin Monoclonal antibody Sensitivity and Specificity complex mixtures Mice Bacterial Proteins Antigen medicine Animals Humans Immunoprecipitation DNA Primers Antigens Bacterial Mice Inbred BALB C CFP-10 biology business.industry Sputum Antibodies Monoclonal Mycobacterium tuberculosis Tuberculosis Pleural bacterial infections and mycoses medicine.disease Virology Disease Models Animal Infectious Diseases Polyclonal antibodies ESAT-6 Monoclonal biology.protein Databases Nucleic Acid business |
| Zdroj: | The International Journal of Tuberculosis and Lung Disease. 15:804-810 |
| ISSN: | 1027-3719 |
| DOI: | 10.5588/ijtld.10.0393 |
| Popis: | Objective To elucidate the potential of monoclonal antibodies (mAbs) of culture filtrate protein 10 (CFP-10) and early secretory antigenic target 6 (ESAT-6) in tuberculosis (TB) diagnosis. Design We generated and characterised monoclonal and polyclonal antibodies against Mycobacterium tuberculosis-specific antigens ESAT-6 and CFP-10 by immunising BALB/c mice with an ESAT-6/CFP-10 fusion protein. Stable hybridoma cell lines were established and mAbs were specifically identified by immunoblotting and immunoprecipitation. The mouse mAbs were used to coat plates, and biotin-labelled polyclonal antibodies were used to detect the antigens. One hundred and seventy-three samples of sputum culture supernatants and pleural effusion aspirates have been tested. Results The ESAT-6 enzyme-linked immunosorbent assay (ELISA) detected the culture supernatants and pleural effusion specimens that were positive for M. tuberculosis, but failed to identify M. tuberculosis-positive specimens in the non-M. tuberculosis culture supernatants or control specimens. This yielded a sensitivity of 95.4% and a specificity of 100% for the ESAT-6-specific ELISA. The CFP-10 ELISA presented less satisfactory sensitivity and specificity, of respectively 81.6% and 92.2%. Results showed positive detection rates of ESAT-6 and CFP-10 of 86.8% (33/38) and 76.3% (29/38) for the diagnosis of tuberculous pleural effusion in patients bacteriologically negative for M. tuberculosis culture. Conclusion The ESAT-6 and CFP-10 ELISAs incorporating mAbs generated in this study serve as potential tools in the laboratory diagnosis of TB. |
| Databáze: | OpenAIRE |
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