title: Identification and characterization of a novel Leishmania donovani antigen for serodiagnosis of visceral leishmaniasis.
abstract: Despite several drawbacks, rK39-based rapid immunochromatographic test is widely used for the diagnosis of visceral leishmaniasis (VL) in the Indian subcontinent. There is an urgent need to develop a better antigen. In this study we separated crude soluble antigens of Leishmania donovani by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and hybridized with pool sera from pre- and post-treated VL patients, 6 months follow-up, endemic healthy (EHC), and nonendemic healthy controls (NEHC) by Western blotting. The sensitivity of enzyme-linked immunosorbent assay with identified protein was 95% (confidence interval [CI] = 89.6-98.01%), whereas the specificity for EHC, NEHC, and different disease groups were 96.3% (CI = 89.8-98.6%), 100% (CI = 95.8-100%), and 97.4% (CI = 91.02-99.3%), respectively. This specific antigen was subjected to two-dimensional gel electrophoresis and after tryptic digestion, antigen was characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Further analysis showed that it is a member of the heat shock protein family of 70 kDa, designated as BHUP1, and has great potential in the diagnosis of VL.
authors.affiliation: Department of Medicine, Institute of Medical Sciences, Banaras Hindu University Varanasi, India. subodh_bt2003@yahoo.co.in
authors.forename: Subodh
authors.initials: S
authors.lastname: Kumar
authors.forename: Dinesh
authors.initials: D
authors.lastname: Kumar
authors.forename: Jaya
authors.initials: J
authors.lastname: Chakravarty
authors.forename: Madhukar
authors.initials: M
authors.lastname: Rai
authors.forename: Shyam
authors.initials: S
authors.lastname: Sundar
mesh_terms: D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000097:blood* / Q000302:isolation & purification; D015153:Blotting, Western / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005500:Follow-Up Studies; D006801:Humans; D007893:Leishmania donovani / Q000276:immunology / Q000302:isolation & purification / Q000472:pathogenicity; D007898:Leishmaniasis, Visceral / Q000175:diagnosis*; D015800:Protozoan Proteins / Q000097:blood / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012698:Serologic Tests
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D015800:Protozoan Proteins