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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/10814
Title: A novel 12.6-kDa protein of leishmania donovani for the diagnosis of Indian visceral leishmaniasis
Authors: Kumar D.
Kumar S.
Chakravarty J.
Sundar S.
Published in: Vector-Borne and Zoonotic Diseases
Abstract: Background: For the diagnosis of visceral leishmaniasis (VL), rK39 antigen-based rapid test is widely used. Unfortunately, up to 32% healthy individuals from endemic region test positive with this antigen. There is an urgent need to search for a more specific antigen with sensitivity similar to rK39. Methods: We identified a Leishmania donovani-specific 12.6-kDa (BHUP3) soluble promastigote antigen through sensitive western blot technique. The identified protein was partially purified from sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the antigenic response of eluted protein was determined by western blot with different groups of individual sera. The diagnostic potential was further validated by enzyme-linked immunosorbent assay using serum of 100 VL patients, 93 nonendemic healthy control individuals, 110 endemic healthy control individuals, and 110 disease control individuals. Further, it was characterized by two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/ionization-time-of-flight analysis. Results: On blotting, antibody against this protein was recognized by all (9/9) VL patient's sera, but it was absent in every control group (nonendemic healthy control and endemic healthy control). Sensitivity of the enzyme-linked immunosorbent assay was 88% (89/101), whereas the specificity for endemic healthy, nonendemic healthy, and different disease groups were 96% (106/110), 100% (93/93), and 97% (107/110), respectively. The two-dimensional gel electrophoresis showed a single spot, and matrix-assisted laser desorption/ionization-time-of-flight analysis revealed that it is a 113-amino-acid-long putative uncharacterized protein of 12.6-kDa anamorsin homolog matched completely with Leishmania major (GenBank accession number: Q4QIS1). Conclusion: Despite marginally lower sensitivity of BHUP3, excellent specificity warrants its further development as a tool for diagnosis of VL. © Copyright 2011, Mary Ann Liebert, Inc.
Citation: Vector-Borne and Zoonotic Diseases (2011), 11(10): 1359-1364
URI: https://doi.org/10.1089/vbz.2011.0620
http://repository.iitr.ac.in/handle/123456789/10814
Issue Date: 2011
Keywords: Diagnosis
ELISA
MALDI-TOF
Visceral leishmaniasis
Western blotting
ISSN: 15303667
Author Scopus IDs: 57202478211
57209548091
16241108100
7103328080
Author Affiliations: Kumar, D., Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, Uttar Pradesh, India
Kumar, S., Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, Uttar Pradesh, India
Chakravarty, J., Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, Uttar Pradesh, India
Sundar, S., Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, Uttar Pradesh, India
Corresponding Author: Sundar, S.; Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, Uttar Pradesh, India; email: drshyamsundar@hotmail.com
Appears in Collections:Journal Publications [ME]

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