Fol. Biol. 2014, 60, 10-20
Efficient ELISA for Diagnosis of Active Tuberculosis Employing a Cocktail of Secretory Proteins of Mycobacterium tuberculosis
Rapid and accurate diagnosis is important for preventing transmission of Mycobacterium tuberculosis. Currently available tuberculosis (TB) diagnostic methods lack desired sensitivity and specificity, and require sophisticated equipment and skilled workforce including weeks’ long duration to yield results. In this study, extracellular proteins or secretory protein antigens of M. tuberculosis H37Rv have been isolated using ion exchange chromatography, immunocharacterized and exploited for the development of efficient enzyme-linked immunosorbent assay (ELISA) for diagnosis of active TB with enhanced specificity and sensitivity. Apparent molecular masses for purified proteins were found to be 6, 27, 30, 38 and 64 kDa. Out of five purified proteins, one (64 kDa) was found to be novel. Of the five proteins, four (6, 27, 30 and 38 kDa) were found significant to be used in the development of ELISA for pulmonary and extra-pulmonary TB. The immune responses of serum samples of TB patients and other healthy subjects against the above-mentioned antigens’ cocktail were evaluated. Critical parameters of newly developed ELISA were optimized and it was observed that the cocktail antigens have a greater specificity (98.06 %) and sensitivity (98.67 %) as compared to other commercially available diagnostic tests. The present findings suggest that the developed ELISA is an effective tool for routine screening and early-stage diagnosis of TB.
Keywords
tuberculosis, culture filtrate proteins, ELISA, MDR, secretory antigens, serodiagnosis.
Funding
The financial support for the research work was jointly provided by the Zydus Research Centre, Ahmedabad, Gujarat, India, Dr. B.R. Ambedkar Centre for Biomedical Research, University of Delhi, Delhi, India and the School of Studies in Biotechnology, Jiwaji University, Gwalior, MP, India.
References
Copyright
This is an open-access article distributed under the terms of the Creative Commons Attribution License.