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Development and evaluation of Leishmania infantum rK26 ELISA for serodiagnosis of visceral leishmaniasis in Iran

Published online by Cambridge University Press:  19 June 2008

S. FARAJNIA ,
B. DARBANI ,
H. BABAEI ,
M. H. ALIMOHAMMADIAN ,
F. MAHBOUDI  and
A. M. GAVGANI
S. FARAJNIA*
Affiliation:
Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
B. DARBANI
Affiliation:
Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
H. BABAEI
Affiliation:
Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran School of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran
M. H. ALIMOHAMMADIAN
Affiliation:
Immunology Department, Pasteur Institute of Iran, Iran
F. MAHBOUDI
Affiliation:
Biotechnology Research Center, Pasteur Institute of Iran, Iran
A. M. GAVGANI
Affiliation:
Infectious and Tropical Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
*
*Corresponding author: Drug Applied Research Center, Tabriz University of Medical Science, Daneshgah Avenue, Tabriz 51656-65811, Iran. Tel: 0098411 3363234. E-Mail: farajnias@tbzmed.ac.ir
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Summary

The purpose of this study was to prepare recombinant K26 antigen from Leishmania infantum and evaluate its performance by enzyme-linked immunosorbent assay (ELISA) test for serodiagnosis of visceral leishmaniasis (VL) in endemic regions of Iran. The results were compared with those obtained by direct agglutination test (DAT) and whole cell ELISA using crude parasite antigen. Of 93 sera from patients with confirmed VL, 90 sera were positive with rK26 ELISA (sensitivity=96·8%), whereas 85 sera were positive with DAT (sensitivity=91·4%) and 89 sera were positive with whole cell ELISA (sensitivity=95·7%). Of 130 subjects who either had other infectious diseases (n=30) or were healthy (n=100), rK26 ELISA were negative in all cases (specificity=100%), whereas DAT were negative in 116 cases (specificity=89·2%) and whole cell ELISA was negative in 114 cases (specificity=87·7%). The results of this study indicate that the rK26 ELISA is more sensitive and specific than conventional methods and could be used for reliable diagnosis of VL caused by Leishmania infantum.

Keywords

Leishmania infantumserodiagnosisrecombinant antigensK26
Type
Original Articles
Information
Parasitology , Volume 135 , Issue 9 , August 2008 , pp. 1035 - 1041
Copyright
Copyright © 2008 Cambridge University Press

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References

REFERENCES

Altman, D. G. (1991). Practical Statistics for Medical Research. Chapman and Hall, London, UK.Google Scholar
Bhatia, A., Daifalla, N. S., Jen, S., Badaro, R., Reed, S. G. and Skeiky, Y. A. (1999). Cloning, characterization and serological evaluation of K9 and K26: two related hydrophilic antigens of Leishmania chagasi. Molecular and Biochemical Parasitology 102, 249–261.CrossRefGoogle ScholarPubMed
Bern, C., Jha, S. N., Joshi, A. B., Thakur, G. D. and Bista, M. B. (2000). Use of the recombinant k39 dipstick test and the direct agglutination test in a setting endemic for visceral leishmaniasis in Nepal. American Journal of Tropical Medicine and Hygiene 63, 153–157.CrossRefGoogle Scholar
Bogdan, C., Stosiek, N., Fuchs, H., Rollinghoff, M. and Solbach, W. (1990). Detection of potentially diagnostic leishmanial antigens by Western blot analysis of sera from patients with kala-azar or multilesional cutaneous leishmaniasis. Journal of Infectious Diseases 162, 1417–1418.CrossRefGoogle ScholarPubMed
Burns, J. M. Jr., Shreffler, W. G., Benson, D. R., Ghalib, H. W., Badaro, R. and Reed, S. G. (1993). Molecular characterization of a kinesin-related antigen of Leishmania chagasi that detects specific antibody in African and American visceral leishmaniasis. Proceedings of the National Academy of Sciences, USA 90, 775–779.CrossRefGoogle ScholarPubMed
Carvalho, F. A., Charest, H., Tavares, C. A., Matlashewski, G., Valente, E. P., Rabello, A., Gazzinelli, R. T. and Fernandes, A. P. (2002). Diagnosis of American visceral leishmaniasis in humans and dogs using the recombinant Leishmania donovani A2 antigen. Diagnostic Microbiology and Infectious Disease 43, 289–295.CrossRefGoogle ScholarPubMed
Chappuis, F., Rijal, S., Soto, A., Menten, J. and Boelaert, M. (2006). A meta-analysis of the diagnostic performance of the direct agglutination test and rK39 dipstick for visceral leishmaniasis. British Medical Journal 333, 723–728.CrossRefGoogle ScholarPubMed
Choudhry, A., Guru, P. Y., Saxena, R. P., Tandon, A. and Saxena, K. C. (1990). Enzyme-linked immunosorbent assay in the diagnosis of kala-azar in Bhadohi (Varanasi), India. Transactions of the Royal Society of Tropical Medicine and Hygiene 84, 363–366.CrossRefGoogle ScholarPubMed
da Costa, R. T., Franca, J. C., Mayrink, W., Nascimento, E., Genaro, O. and Campos-Neto, A. (2003). Standardization of a rapid immunochromatographic test with the recombinant antigens K39 and K26 for the diagnosis of canine visceral leishmaniasis. Transactions of the Royal Society of Tropical Medicine and Hygiene 97, 678–682.CrossRefGoogle ScholarPubMed
D'Oliveira, J. A., Costa, S. R., Barbosa, A. B., Orge, M. G. and Carvalho, E. M. (1997). Asymptomatic Leishmania chagasi infection in relatives and neighbors of patients with visceral leishmaniasis. Memórias do Instituto Oswaldo Cruz 92, 15–20.CrossRefGoogle Scholar
el Harith, A., Kolk, A. H., Leeuwenburg, J., Muigai, R., Huigen, E., Jelsma, T. and Kager, P. A. (1998). Improvement of a direct agglutination test for field studies of visceral leishmaniasis. Journal of Clinical Microbiology 26, 1321–1325.CrossRefGoogle Scholar
Farajnia, S., Alimohammadian, M. H., Reiner, N. E., Karimi, M., Ajdari, S. and Mahboudi, F. (2004). Molecular characterization of a novel amastigote stage specific Class I nuclease from Leishmania major. International Journal for Parasitology 34, 899–908.CrossRefGoogle Scholar
Farajnia, S., Mahboudi, F., Ajdari, S., Reiner, N. E., Kariminia, A. and Alimohammadian, M. H. (2005). Mononuclear cells from patients recovered from cutaneous leishmaniasis respond to Leishmania major amastigote class I nuclease with a predominant Th1-like response. Clinical and Experimental Immunology 139, 498–505.CrossRefGoogle Scholar
Guerin, P. J., Olliaro, P., Sundar, S., Boelart, M., Croft, S. L., Desjeux, P., Wasunna, M. K. and Bryceson, A. (2002). Visceral leishmaniasis: current status of control, diagnosis and treatment, and a proposed research and development agenda. Lancet Infectious Disease 2, 494–501.CrossRefGoogle Scholar
Jelinek, T., Eichenlaub, S. and Löscher, T. (1999). Sensitivity and specificity of a rapid immunochromatographic test for diagnosis of visceral leishmaniasis. European Journal of Clinical Microbiology and Infectious Disease 18, 669–670.CrossRefGoogle ScholarPubMed
Kar, K. (1995). Serodiagnosis of leishmaniasis. Critical Revews in Microbiology 21, 123–152.CrossRefGoogle ScholarPubMed
Kumar, R., Pai, K., Pathak, K. and Sundar, S. (2001). Enzyme-linked immunosorbent assay for recombinant K39 antigen in diagnosis and prognosis of Indian visceral leishmaniasis. Clinical and Diagnostic Laboratory Immunology 8, 1220–1224.CrossRefGoogle ScholarPubMed
Lowry, O. H., Roseborough, N. J., Farr, A. L. and Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 139, 265–275.CrossRefGoogle Scholar
Maalej, I. A., Chenik, M., Louzir, H., Ben Salah, A., Bahloul, C., Amri, F. and Dellagi, K. (2003). Comparative evaluation of ELISA's based on ten recombinant or purified Leishmania antigens for the serodiagnosis of mediterranean visceral leishmaniasis. American Journal of Tropical Medicine and Hygiene 68, 312–320.CrossRefGoogle ScholarPubMed
Malla, N., Sengupta, C., Dubey, M. L., Sud, A., Ansari, N. A. and Salotra, P. (2003). Antigenaemia and antibody response to Leishmania donovani stage-specific antigens and rk39 antigen in human immunodeficiency virus-infected patients. British Journal of Biomedical Science 60, 210–216.CrossRefGoogle ScholarPubMed
Millesimo, M., Zucca, M., Caramello, P. and Savoia, D. (1996). Evaluation of the immune response in visceral leishmaniasis. Diagnostic Microbiology and Infectious Disease 26, 7–11.CrossRefGoogle ScholarPubMed
Mohebali, M., Taran, M. and Zarei, Z. (2004). Rapid detection of Leishmania infantum infection in dogs: comparative study using an immunochromatographic dipstick rk39 test and direct agglutination. Veterinary Parasitology 26, 239–245.CrossRefGoogle Scholar
Pal, A., Mukerji, K., Basu, D., Naskar, K., Mallick, K. K. and Ghosh, D. K. (1991). Evaluation of a direct agglutination test (DAT) and ELISA for serodiagnosis of visceral leishmaniasis in India. Journal of Clinical Laboratory Analysis 5, 303–306.CrossRefGoogle ScholarPubMed
Rosário, E. Y., Genaro, O., Franca-Silva, J. C., da Costa, R. T., Mayrink, W., Reis, A. B. and Carneiro, M. (2005). Evaluation of enzyme-linked immunosorbent assay using crude Leishmania and recombinant antigens as a diagnostic marker for canine visceral leishmaniasis. Memórias do Instituto Oswaldo Cruz 100, 197–203.CrossRefGoogle ScholarPubMed
Rosati, S., Ortoffi, M., Profiti, M., Mannelli, A., Mignone, W., Bollo, E. and Gradoni, L. (2003). Prokaryotic expression and antigenic characterization of three recombinant Leishmania antigens for serological diagnosis of canine leishmaniasis. Clinical and Diagnostic Laboratory Immunology 10, 1153–1156.Google ScholarPubMed
Singh, S., Gilman-Sachs, A., Chang, K. P. and Reed, S. G. (1995). Diagnostic and prognostic value of K39 recombinant antigen in Indian leishmaniasis. Journal of Parasitology 81, 1000–1003.CrossRefGoogle ScholarPubMed
Sivakumar, R., Sharma, P., Chang, K. P. and Singh, S. (2006). Cloning, expression, and purification of a novel recombinant antigen from Leishmania donovani. Protein Expression and Purification 46, 156–165.CrossRefGoogle ScholarPubMed
Sundar, S., Pai, K., Sahu, M., Kumar, V. and Murray, H. W. (2002). Immunochromatogaphic strip test detection of anti k39 antibody in Indian visceral leishmaniasis. Annals of Tropical Medicine and Parasitology 96, 19–23.CrossRefGoogle Scholar
Sundar, S. and Rai, M. (2002). Laboratory diagnosis of visceral leishmaniasis. Clinical and Diagnostic Laboratory Immunology 9, 951–958.Google ScholarPubMed
Sundar, S., Singh, R. K., Bimal, S. K., Gidwani, K., Mishra, A., Maurya, R., Singh, S. K., Manandhar, K. D., Boelaert, M. and Rai, M. (2007). Comparative evaluation of parasitology and serological tests in the diagnosis of visceral leishmaniasis in India: a phase III diagnostic accuracy study. Tropical Medicine and International Health 12, 284–289.CrossRefGoogle ScholarPubMed
Towbin, H., Staehelin, T. and Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, USA 76, 4350–4354.CrossRefGoogle ScholarPubMed
Zijlstra, E. E., Ali, M. S., el-Hassan, A. M., el-Toum, I. A., Satti, M., Ghalib, H. W. and Kager, P. A. (1991). Direct agglutination test for diagnosis and seroepidemiological survey of kala-azar in the Sudan. Transactions of the Royal Society of Tropical Medicine and Hygiene 85, 474–476.CrossRefGoogle ScholarPubMed