Volume 86, Issue 4
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



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 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.


Article metrics loading...

The graphs shown below represent data from March 2017
Loading full text...

Full text loading...



  1. Harith AE, Kolk AH, Kager PA, Leeuwenburg J, Muigai R, Kiugu S, Laarman JJ, , 1986. A simple and economical direct agglutination test for serodiagnosis and sero-epidemiological studies of visceral leishmaniasis. Trans R Soc Trop Med Hyg 80: 583636.[Crossref] [Google Scholar]
  2. el Harith A, Kolk AH, Leeuwenburg J, Muigai R, Huigen E, Jelsma T, Kager PA, , 1988. Improvement of a direct agglutination test for field studies of visceral leishmaniasis. J Clin Microbiol 26: 13211325. [Google Scholar]
  3. Boelaert M, El-Safi S, Hailu A, Mukhtar M, Rijal S, Sundar S, Wasunna M, Aseffa A, Mbui J, Menten J, Desjeux P, Peeling RW, , 2008. Diagnostic tests for kala-azar: a multi-centre study of the freeze-dried DAT, rK39 strip test and KAtex in East Africa and the Indian subcontinent. Trans R Soc Trop Med Hyg 102: 3240.[Crossref] [Google Scholar]
  4. Burns JM, Jr Shreffler WG, Benson DR, Ghalib HW, Badaro R, Reed SG, , 1993. Molecular characterization of a kinesin-related antigen of Leishmania chagasi that detects specific antibody in African and American visceral leishmaniasis. Proc Natl Acad Sci USA 90: 775779.[Crossref] [Google Scholar]
  5. Zijlstra EE, Daifalla NS, Kager PA, Khalil EA, El-Hassan AM, Reed SG, , Ghalib HW, 1998. rK39 enzyme-linked immunosorbent assay for diagnosis of Leishmania donovani infection. Clin Diagn Lab Immunol 5: 717720. [Google Scholar]
  6. Sundar S, Reed SG, Singh VP, Kumar PC, Murray HW, , 1998. Rapid accurate field diagnosis of Indian visceral leishmaniasis. Lancet 351: 563565.[Crossref] [Google Scholar]
  7. Sundar S, Sahu M, Mehta H, Gupta A, Kohli U, Rai M, Berman JD, Murray HW, , 2002. Noninvasive management of Indian visceral leishmaniasis: clinical application of diagnosis by K39 antigen strip testing at a kala-azar referral unit. Clin Infect Dis 35: 581586.[Crossref] [Google Scholar]
  8. Sundar S, Singh RK, Maurya R, Kumar B, Chhabra A, Singh V, Rai M, , 2006. Serological diagnosis of Indian visceral leishmaniasis: direct agglutination test versus rK39 strip test. Trans R Soc Trop Med Hyg 100: 533537.[Crossref] [Google Scholar]
  9. Laemmli VK, , 1970. Cleavage of structural protein during the assembly of the head of baceriophase T4. Nature 227: 682685. [Google Scholar]
  10. Towbin HS, Staehelin T, Gordon J, , 1979. Electrophoretic transfer of polyacrylamide gels to nitrocellulose sheet: procedure and some applications. Proc Natl Acad Sci USA 76: 43504354.[Crossref] [Google Scholar]
  11. Hommel M, Peters W, Ranque J, Quilici M, Lanotte G, , 1978. The micro-ELISA technique in the serodiagnosis of visceral leishmaniasis. Ann Trop Med Parasitol 72: 213218.[Crossref] [Google Scholar]
  12. MacFarlane J, Blaxter ML, Bishop RP, Miles MA, Kelly JM, , 1990. Identification and characterization of a Leishmania donovani antigen belonging to the 70-kDa heat-shock protein family. Eur J Biochem 190: 377384.[Crossref] [Google Scholar]
  13. Gidwani K, Picado A, Ostyn B, Singh SP, Kumar R, Khanal B, Lejon V, Chappuis F, Boelaert M, Sundar S, , 2011. Persistence of Leishmania donovani antibodies in past visceral leishmaniasis cases in India. Clin Vaccine Immunol 18: 346348.[Crossref] [Google Scholar]
  14. Rico AI, Girones N, Fresno M, Alonso C, Requena JM, , 2002. The heat shock proteins, Hsp70 and Hsp83, of Leishmania infantum are mitogens for mouse B cells. Cell Stress Chaperones 7: 339346.[Crossref] [Google Scholar]
  15. Wallace GR, Ball AE, MacFarlane J, el Safi SH, Miles MA, Kelly JM, , 1992. Mapping of a visceral leishmaniasis-specific immunodominant B-cell epitope of Leishmania donovani Hsp70. Infect Immun 60: 26882693. [Google Scholar]
  16. de Andrade CR, Kirchhoff LV, Donelson JE, Otsu K, , 1992. Recombinant Leishmania Hsp90 and Hsp70 are recognized by sera from visceral leishmaniasis patients but not Chagas’ disease patients. J Clin Microbiol 30: 330335. [Google Scholar]
  17. Rafati S, Gholami E, Hassani N, Ghaemimanesh F, Taslimi Y, Taheri T, Soong L, , 2007. Leishmania major heat shock protein 70 (HSP70) is not protective in murine models of cutaneous leishmaniasis and stimulates strong humoral responses in cutaneous and visceral leishmaniasis patients. Vaccine 25: 41594169.[Crossref] [Google Scholar]
  18. Quijada L, Requena JM, Soto M, Alonso C, , 1998. Analysis of the antigenic properties of the L. infantum Hsp70: design of synthetic peptides for specific serodiagnosis of human leishmaniasis. Immunol Lett 63: 169174.[Crossref] [Google Scholar]

Data & Media loading...

  • Received : 25 Apr 2011
  • Accepted : 05 Jul 2011
  • Published online : 01 Apr 2012

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error