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-
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 : 583– 636.
-
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 : 1321– 1325.
-
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 : 32– 40.
-
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 : 775– 779.
-
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 : 717– 720.
-
6.
Sundar S, Reed SG, Singh VP, Kumar PC, Murray HW, 1998. Rapid accurate field diagnosis of Indian visceral leishmaniasis. Lancet 351 : 563– 565.
-
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 : 581– 586.
-
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 : 533– 537.
-
9.
Laemmli VK, 1970. Cleavage of structural protein during the assembly of the head of baceriophase T4. Nature 227 : 682– 685.
-
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 : 4350– 4354.
-
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 : 213– 218.
-
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 : 377– 384.
-
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 : 346– 348.
-
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 : 339– 346.
-
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 : 2688– 2693.
-
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 : 330– 335.
-
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 : 4159– 4169.
-
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 : 169– 174.
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Identification and Characterization of a Novel Leishmania donovani Antigen for Serodiagnosis of Visceral Leishmaniasis
Subodh Kumar
Subodh Kumar
Department of Medicine, Institute of Medical Sciences, Banaras Hindu University Varanasi–India
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Dinesh Kumar
Dinesh Kumar
Department of Medicine, Institute of Medical Sciences, Banaras Hindu University Varanasi–India
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Jaya Chakravarty
Jaya Chakravarty
Department of Medicine, Institute of Medical Sciences, Banaras Hindu University Varanasi–India
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Madhukar Rai
Madhukar Rai
Department of Medicine, Institute of Medical Sciences, Banaras Hindu University Varanasi–India
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Shyam Sundar
Shyam Sundar
Department of Medicine, Institute of Medical Sciences, Banaras Hindu University Varanasi–India
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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.
Author Notes
Financial support: This work was supported by NIAID, NIH grant no.: 1P50AI074321. Subodh Kumar and Dinesh Kumar thank the Council of Scientific and Industrial Research (CSIR), New Delhi, India for providing fellowship.
Authors’ addresses: Subodh Kumar, Dinesh Kumar, Jaya Chakravarty, Madhukar Rai, and Shyam Sundar, Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, E-mails: subodh_bt2003@yahoo.co.in, praja_dinesh@yahoo.co.in, tapadar@gmail.com, rai_madhukar@gmail.com, and drshyamsundar@hotmail.com.
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
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 : 583– 636.
-
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 : 1321– 1325.
-
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 : 32– 40.
-
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 : 775– 779.
-
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 : 717– 720.
-
6.
Sundar S, Reed SG, Singh VP, Kumar PC, Murray HW, 1998. Rapid accurate field diagnosis of Indian visceral leishmaniasis. Lancet 351 : 563– 565.
-
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 : 581– 586.
-
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 : 533– 537.
-
9.
Laemmli VK, 1970. Cleavage of structural protein during the assembly of the head of baceriophase T4. Nature 227 : 682– 685.
-
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 : 4350– 4354.
-
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 : 213– 218.
-
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 : 377– 384.
-
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 : 346– 348.
-
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 : 339– 346.
-
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 : 2688– 2693.
-
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 : 330– 335.
-
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 : 4159– 4169.
-
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 : 169– 174.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 154 | 134 | 10 |
| Full Text Views | 1127 | 9 | 0 |
| PDF Downloads | 74 | 2 | 0 |