Novel monoclonal antibodies to ESAT-6 and CFP-10 antigens for ELISA-based diagnosis of pleural tuberculosis.

Abstract

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: T-T Feng, C-M Shou, L Shen, Y Qian, Z-G Wu, J Fan, Y-Z Zhang, Y-W Tang, N-P Wu, H-Z Lu, H-P Yao
• Journal: The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease (Int J Tuberc Lung Dis) Vol. 15 Issue 6 Pg. 804-10 (Jun 2011) ISSN: 1815-7920 [Electronic] France
• PMID: 21575303 (Publication Type: Comparative Study, Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• Antigens, Bacterial
• Bacterial Proteins
• CFP-10 protein, Mycobacterium tuberculosis
• DNA Primers
• ESAT-6 protein, Mycobacterium tuberculosis
• Tuberculin

Topics

• Animals
• Antibodies, Monoclonal
• Antigens, Bacterial (immunology)
• Bacterial Proteins (immunology)
• DNA Primers
• Databases, Nucleic Acid
• Disease Models, Animal
• Enzyme-Linked Immunosorbent Assay (methods)
• Humans
• Immunoblotting
• Immunoprecipitation
• Mice
• Mice, Inbred BALB C
• Mycobacterium tuberculosis (immunology, isolation & purification)
• Sensitivity and Specificity
• Sputum (microbiology)
• Tuberculin (immunology)
• Tuberculosis, Pleural (diagnosis)