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.
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.
Authors: Chang Liu; Zhen Zhao; Jia Fan; Christopher J Lyon; Hung-Jen Wu; Dobrin Nedelkov; Adrian M Zelazny; Kenneth N Olivier; Lisa H Cazares; Steven M Holland; Edward A Graviss; Ye Hu Journal: Proc Natl Acad Sci U S A Date: 2017-03-27 Impact factor: 11.205
Authors: Zaida Araujo; Francesca Giampietro; María de los Angeles Bochichio; Andrea Palacios; Jenifer Dinis; Jaime Isern; Jacobus Henry de Waard; Elsa Rada; Rafael Borges; Carlos Fernández de Larrea; Angel Villasmil; Magnolia Vanegas; Jose Antonio Enciso-Moreno; Manuel Alfonso Patarroyo Journal: Mem Inst Oswaldo Cruz Date: 2013-04 Impact factor: 2.743
Authors: Chang Liu; Christopher J Lyon; Yang Bu; Zaian Deng; Elisabetta Walters; Yan Li; Liqun Zhang; Anneke C Hesseling; Edward A Graviss; Ye Hu Journal: Clin Chem Date: 2018-01-18 Impact factor: 8.327
Authors: Jia Fan; Hedong Zhang; Duc T Nguyen; Christopher J Lyon; Charles D Mitchell; Zhen Zhao; Edward A Graviss; Ye Hu Journal: BMC Med Date: 2017-11-01 Impact factor: 8.775
Authors: Dewi Kartika Turbawaty; Adhi Kristianto Sugianli; Arto Yuwono Soeroto; Budi Setiabudiawan; Ida Parwati Journal: Int J Microbiol Date: 2017-10-18