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Highly immunoreactive antibodies against the rHup-F2 fragment (aa 63–161) of the iron-regulated HupB protein of Mycobacterium tuberculosis and its potential for the serodiagnosis of extrapulmonary and recurrent tuberculosis

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Abstract

HupB is an iron-regulated protein in Mycobacterium tuberculosis that functions as a positive regulator of mycobactin biosynthesis. It is essential for the growth and survival of the pathogen inside macrophages. Previously, using the full-length rHupB of M. tuberculosis, we demonstrated high levels of anti-HupB antibodies in the serum of pulmonary tuberculosis (TB) and, interestingly, extrapulmonary TB patients with negligible levels in household contacts and healthy controls. Here, we used three antigenic fragments of HupB, namely the recombinant HupB-F1 (aa 1–71), HupB-F2 (aa 63–161) and HupB-F3 (aa 164–214), as antigens in enzyme-linked immunosorbent assay (ELISA) to screen serum from TB patients. HupB-F2 showed enhanced immunoreactivity with serum from patients with pulmonary TB (three groups consisting of new cases, defaulters and recurrent cases) and extrapulmonary TB, with negligible levels in normal healthy controls. The negative correlation of the anti-(HupB-F2) antibodies with serum iron was maximal, with a Pearson’s correlation coefficient value of −0.415. The study, in addition to strengthening the diagnostic potential of HupB, reflected the superior performance of HupB-F2 as an antigen in screening pulmonary and extrapulmonary TB.

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Acknowledgements

MC, SS and NC thank the University Grants Commission for financial assistance as Senior Research Fellows. NS acknowledges the Indian Council of Medical Research for awarding a Short Term Research Fellowship for this study and thanks NRI Medical College for allowing her to carry out the study. MS would like to acknowledge the Council of Scientific and Industrial Research (CSIR) for the financial assistance for performing part of the study. We would like to thank Ms. Kiranmayi Vemuri for her help in the purification of recombinant HupB-F3 protein.

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Authors and Affiliations

  1. Department of Animal Sciences, School of Life Sciences, University of Hyderabad, Prof. CR Rao Road, Gachibowli, Hyderabad, 500 046, India

    M. Choudhury, S. Sivakolundu, R. Chaurasia, N. Chouhan & M. Sritharan

  2. NRI Academy of Sciences, Guntur, Andhra Pradesh, India

    N. Sritharan & P. P. Rao

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  1. N. Sritharan
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  2. M. Choudhury
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  3. S. Sivakolundu
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  4. R. Chaurasia
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  5. N. Chouhan
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  6. P. P. Rao
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  7. M. Sritharan
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Correspondence to M. Sritharan.

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Sritharan, N., Choudhury, M., Sivakolundu, S. et al. Highly immunoreactive antibodies against the rHup-F2 fragment (aa 63–161) of the iron-regulated HupB protein of Mycobacterium tuberculosis and its potential for the serodiagnosis of extrapulmonary and recurrent tuberculosis. Eur J Clin Microbiol Infect Dis 34, 33–40 (2015). https://doi.org/10.1007/s10096-014-2203-y

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  • DOI: https://doi.org/10.1007/s10096-014-2203-y

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