Volume 68, Issue 3
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645


This study compared a panel of 10 enzyme-linked immunosorbent assays (ELISAs) for the serodiagnosis of Mediterranean visceral leishmaniasis (MVL). The ELISAs were based on either one of the following antigens: crude soluble antigens (SLAs), recombinant (r) antigens (namely: rgp63, rK39, gene B protein, rH2A and rH2B histones proteins, rLACK, rPSA-2, r P20) and purified lipophosphoglycan. Most of the test antigens showed good performance (sensitivity > 85%, specificity > 80%). rK39 and SLA-based ELISA gave the best results in terms of sensitivity (100%) and predictive value of the negative (100%). The best specificity (97%) and the best predictive value of the positive (92%) were obtained with rK39. These results show that several antigens are suitable to design a diagnostic ELISA of MVL. However, recombinant proteins add little to the classic crude SLA, which still represents a very good and less costly alternative.


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  1. Peters W, Killick-Kendrick R, 1987. The Leishmaniases in Biology and Medicine. London: Academic Press.
  2. Ben-Ismail R, Ben Rachid MS, 1989. Epidémiologie des leishmanioses en Tunisie. AUPELF-UREF. Maladies Tropicales Transmissibles. Paris: John Libbey Eurotext, 73–80.
  3. Momen H, Grimaldi G Jr, Deane LM, 1987. Leishmania infantum, the oetiological agent of American Visceral Leishmaniasis. Mem Inst Oswaldo Cruz 82 : 667–668.
  4. Marty P, Le Fichoux Y, Pratlong F, Gari-Toussaint M, 1994. human visceral leishmaniasis in Alpes-Maritimes, France: epidemiological characteristics for the period 1985–1992. Trans R Soc Trop Med Hyg 47: 764–771.
  5. Badaro R, Reed SG, Carvalho EM, 1983. Immunofluorescence test in American visceral leishmaniasis: sensitivity and specificity of different morphological forms of two Leishmania species. Am J Trop Med Hyg 32 : 480–484.
  6. Basu D, Mallik KK, 1992. Fluorescence antibody titers after recovery from visceral leishmaniasis. Acta Trop 50 : 87–88.
  7. Rolland-Burger L, Rolland X, Grieve CW, Monjour L, 1991. Immunoblot analysis of the humoral immune response to Leishmania donovani infantum polypeptides in human visceral leishmaniasis. J Clin Microbiol 29 : 1429–1435.
  8. Zijlstra EE, Osman OF, Hofland HW, Oskam L, Ghalib HW, El-Hassan AM, Kager PA, Meredith SE, 1997. The direct agglutination test for diagnosis of visceral leishmaniasis under field conditions in Sudan: comparison of aqueous and freeze-dried antigens. Trans R Soc Trop Med Hyg 91 : 671–673.
  9. Badaro R, Reed SG, Barral A, Orge G, Jones TC, 1986. Evaluation of the micro enzyme-linked immunosorbent assay (ELISA) for antibodies in American visceral leishmaniasis: antigen selection for detection of infection-specific responses. Am J Trop Med Hyg 35: 72–78.
  10. Badaro R, Benson D, Eulalio MC, Freire M, Cunha S, Netto EM, Pedral-Sampaio D, Madureira D, Burns JM, Houghton RL, David JR, Reed SG, 1996. rK39: a cloned antigen of Leishmania chagasi that predicts active visceral leishmaniasis. J Infect Dis 173 : 758–761.
  11. 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 U S A 90 : 775–779.
  12. Ho M, Leeuwenburg J, Mbungua G, Wamachi A, Voller A, 1983. An enzyme-linked immunosorbent assay (ELISA) for field diagnosis of visceral leishmaniasis. Am J Trop Med Hyg 32 : 943–946.
  13. Okong’o-odera EA, Kurtzhals JA, Hey AD, Kharazmi A, 1993. Measurement of serum antibodies against native Leishmania gp63 distinguishes between ongoing and previous L. donovani infection. APMIS 101 : 642–646.
  14. Shreffler WG, Burns JM, Badaro R, Ghalib HW, Button LL, McMaster WR, Reed SG, 1993. Antibody responses of visceral leishmaniasis patients to gp63, a major surface glycoprotein of Leishmania species. J Infect Dis 167: 426–430.
  15. Tebourski F, El Gaied A, Louzir H, Ben Ismail R, Kammoun R, Dellagi K, 1994. Identification of an immunodominant 32-Kilodalton membrane protein of Leishmania donovani infantum promastigotes suitable for specific diagnosis of Mediterranean visceral leishmaniasis. J Clin Microbiol 32 : 2474–2480.
  16. Flinn HM, Rangarajan D, Smith DH, 1994. Expression of a hydrophilic surface protein in infective stages of Leishmania major. Mol Biochem Parasitol 65 : 259–270.
  17. Soto M, Requena JM, Gomez LC, Navarete I, Alonso C, 1992. Molecular characterization of a Leishmania donovani infantum antigen identified as histone H2A. Eur J Biochem 205 : 211–216.
  18. Soto M, Requena JM, Quijada Garcia M, Guzman F, Patarroyo ME, Alonso C, 1995. Mapping of the linear antigenic determinants from the Leishmania infantum histone H2A recognized by sera from dogs with leishmaniasis. Immunol Lett 48 : 209–214.
  19. Soto M, Requena JM, Quijada L, Perez MJ, Nieto CG, Guzman F, Patarroyo ME, Alonso C, 1999. Antigenicity of the Leishmania infantum histones H2B and H4 during canine viscerocutaneous leishmaniasis. Clin Exp Immunol 115 : 342–349.
  20. Mougneau E, Altara F, Wakil AE, Zheng S, Coppola T, Wang ZE, Waldmann R, Locksley RM, Glaichenhaus N, 1995. Expression cloning of a protective Leishmania antigen. Science 268 : 563–566.
  21. Handman E, Osborn AH, Symons F, Van Driel R, Cappai R, 1995. The Leishmania promastigote surface antigen 2 complex is differentially expressed during the parasite life cycle. Mol Biochem Parasitol 74 : 189–200.
  22. Murray PJ, Spithill TW, 1991. Variants of a Leishmania surface antigen derived from a multigenic family. J Biol Chem 266 : 24477–24484.
  23. Turco SJ, 1988. The lipophosphoglycan of Leishmania. Parasitol Today 4 : 255–257.
  24. Ayadi A, Ben-Ismail R, Ben-Rachid MS, 1991. Extension de l’aire de transmission du kala-azar à Leishmania infantum (Nicolle 1908) vers le centre et le sud de la Tunisie. Arch Inst Pasteur Tunis 68 : 269–273.
  25. Ben Salah A, Ben Ismail R, Amri F, Chlif S, Ben Rzig F, Kharrat H, Hadhri H, Hassouna M, Dellagi K, 2000. Investigation of the spread of human visceral leishmaniasis in central Tunisia. Trans R Soc Trop Med Hyg 94: 382–386.
  26. Scott P, Pearce E, Natovitz P, Sher A, 1987. Vaccination against cutaneous leishmaniasis in a murine model. I. Induction of protective immunity with a soluble extract of promastigotes. J Immunol 139: 221–227.
  27. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ, 1951. Protein measurement with the Folin phenol reagent. J Biol Chem 37 : 171–182.
  28. Jensen ATR, Gaafar A, Ismail A, Christensen CBV, Kemp M, El Hassan AM, Kharazmi A, Theander TG, 1996. Serodiagnosis of cutaneous leishmaniasis: assessement of an enzyme-linked immunosorbent assay using a peptide sequence from gene B protein. Am J Trop Med Hyg 55 : 490–495.
  29. McKean PG, Delahay R, Pimenta PFP, Smith DF, 1997. Characterization of a second protein encoded by differentially regulated Lmc DNA 16 gene family of Leishmania major. Mol Biochem Parasitol 85 : 221–231.
  30. McKean PG, Trenholme KR, Rangarajan D, Keen JK, Smith DF, 1997. Diversity in repeat-containing surface proteins of Leishmania major. Mol Biochem Parasitol 86 : 225–235
  31. Bates PA, 1995. The lipophosphoglycan-associated molecules of Leishmania. Parasitol Today 9: 317–318.
  32. Julia V, Rassoulzadegan M, Glaichenhaus N, 1996. Resistance to Leishmania major induced by tolerance to a single antigen. Science 274: 421–423.
  33. Russo DM, Turco SJ, Burns JM, Reed SG, 1992. Stimulation of human T lymphocytes by Leishmania lipophosphoglycan-associated proteins. J Immunol 148 : 202–207.
  34. Galanti N, Galindo M, Sabaj V, Espinoza I, Toro GC, 1998. Histone genes in trypanosomatids. Parasitol Today 14 : 64–70.
  35. Gurunathan BS, Sacks DL, Brown DR, Reiner SL, Charest H, Glaichenhaus N, Seder RA, 1997. Vaccination with DNA encoding the immunodominant LACK parasite antigen confers protective immunity to mice infected with Leishmania major. J Exp Med 186: 1137–1147.
  36. Julia V, Glaichenhaus N, 1999. CD4+ T cells which react to Leishmania major LACK antigen rapidly secrete interleukin-4 and are detrimental to the host in resistant B10.D2 mice. Infect Immun 67 : 3641–3644.
  37. Locksley RM, Pingel S, Lacy D, Wakil AE, Bix M, Fowell DJ, 1999. Susceptibility to infectious diseases: Leishmania as a paradigm. J Inf Dis 179 : 305–308.
  38. Daniel TM, Debanne S, 1987. The serodiagnosis of tuberculosis and other mycobacterial diseases by enzyme-linked immunosorbent assay. Am Rev Respir Dis 135 : 1137–1151.
  39. Galen RS, 1982. Application of the predictive value model in the analysis of the test effectiveness.Clin Lab Med 2 : 685–699.
  40. Badaro R, Jones TC, Carvalho EM, Sampaio D, Reed SG, Barral A, Teixeira R, Johnson WD Jr, 1986. New prospectives on a subclinical form of visceral leishmaniasis. J Infect Dis 154 : 1003–1011.
  41. Evans TG, Teixeira MJ, Sousa AQ, Pearson RD, 1995. Short report: extended follow-up of the natural history of persons infected with Leishmania chagasi. Am J Trop Med Hyg 53:360–361.
  42. Evans TG, Teixeira MJ, McAuliffe IT, Vasconcelos IAB, Vasconcelos AW, Sousa AQ, Lima JWO, Pearson RD, 1992. Epidemiology of visceral leishmaniasis in Northeast Brazil. J Infect Dis 166 : 1124–1132.
  43. Santos ID, Morgado MG, Galvao-Castro B, 1987. Human visceral leishmaniasis: analysis of the specificity of humoral immune response to polypeptides of Leishmania donovani chagasi. Am J Trop Med Hyg 37 : 263–270.
  44. Ozensoy S, Ozbel Y, Turgay N, Alkan MZ, Gul K, Gilman-Sachs A, Chang K-P, Reed SG Ozcel MA, 1998. Serodiagnosis and epidemiology of visceral leishmaniasis in Turkey. Am J Trop Med Hyg 59 : 363–369.
  45. Sundar S, Reed SG, Singh VP, Kumar PCK, Murry HW, 1998. Rapid and accurate field diagnosis of Indian visceral leishmaniasis. Lancet 351 : 563–565.
  46. Jensen ATR, Gasim S, Moller T, Ismail A, Gaafar A, Kemp M, El Hassan AM, Kharazmi A, Alce TM, Smith DF, Theander TG, 1999. Serodiagnosis of Leishmania donovani infection: assessment of enzyme-linked immunosorbent assays using recombinant L. donovani gene B protein (GBP) and a peptide sequence of L. donovani GBP. Trans R Soc Trop Med Hyg 93 : 157–160.
  47. Kurtzhals JAL, Hey AS, Theander TG, Odera E, Christensen CBV, Githure JL, Koech DK, Schaefer KU, Handman E, Kharazmi A, 1992. Cellular and humoral immune response in a population from the Baringo district, Kenya, to Leishmania promastigote lipophosphoglycan. Am J Trop Med Hyg 46 : 480–488.

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  • Received : 26 Feb 2002
  • Accepted : 12 Aug 2002

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