Western Blot Analysis of Leishmania infantum Antigens in Se-ra of Patients with Visceral Leishmaniasis
AbstractBackground: Visceral leishmaniasis (VL) is endemic in the northwest and south of Iran. Untreated cases of VL could cause death. The aim of the present study was to evaluate the diagnostic performance of western blotting to detect a specific immunodominant proteins pattern for Leishmania infantum infection using human sera infected with VL. Methods: We studied a panel of 122 cryopreserved human serum samples from the leishmaniasis Research Laboratory, Tehran University of Medical Sciences, Tehran, Iran from 2010 to 2017. Serum samples were collected from visceral (Group I, n: 43) and cutaneous leishmaniasis (CL) (Group II, n: 8) patients, healthy individuals from endemic (Group III, n: 13) and non-endemic (Group IV, n: 16) areas for VL, and patients with other infectious diseases (Group V, n: 42). Total antigens were prepared from the Iranian strain of L. infantum promastigote form. Results: In western blotting method, 34 protein bands of 14 to 163 kDa were recognized using the sera of VL patients. The polypeptide fractions with the highest frequency including 29, 51, and 62 kDa fractions were detected using 81.4%, 79%, and 81.4% of the sera, respectively. These bands were not detected using the sera of the negative control. Moreover, 19-23, 27, 31-35, 143-163, and 109 kDa fractions were detected specifically using the sera of the patients with VL. Conclusion: This technique could be a primary step for further exploration of VL immunodominant antigens for cloning (or any technique) further investigations for future planning.
Alvar J, Vélez ID, Bern C et al. Leishmaniasis worldwide and global estimates of its incidence. PloS one. 2012;7(5):e35671.
World Health Organization. Report of a meeting of the WHO Expert Committee on the Control of Leishmaniases, Geneva 22-26 March 2010.
Shirzadi M, Esfahania S, Mohebalia M et al. Epidemiological status of leishmaniasis in the Islamic Republic of Iran, 1983-2012. East Mediterr Health J. 2015;21(10):736.
Hajjaran H, Mohebali M, Teimouri A et al. Identification and phylogenetic relationship of Iranian strains of various Leishmania species isolated from cutaneous and visceral cases of leishmaniasis based on N-acetylglucosamine-1-phosphate transferase gene. Infect Genet Evol. 2014;26:203-212.
Mohebali M. Epidemiological status of visceral leishmaniasis in Iran: experiences and review of literature. J Clin Exp Pathol. 2012;S3:003.
Tanoli ZM, Rai ME, Gandapur A. Clinical presentation and management of visceral leishmaniasis. J Ayub Med Coll Abbottabad. 2005;17(4):51-53.
Boelaert M, Verdonck K, Menten J et al. Rapid tests for the diagnosis of visceral leishmaniasis in patients with suspected disease. Cochrane Database Syst Rev. 2014;20:1–119.
De Ruiter C, Van der Veer C, Leeflang M et al. Molecular tools for diagnosis of visceral leishmaniasis: systematic review and meta-analysis of diagnostic test accuracy. J clin microbiol. 14;52(9):3147-3155.
Boelaert M, Rijal S, Regmi S et al. A comparative study of the effectiveness of diagnostic tests for visceral leishmaniasis. Am J Trop Med Hyg. 2004;70(1):72-77.
Elmahallawy EK, Martinez AS, Rodriguez-Granger J et al. Diagnosis of leishmaniasis. J Infect Dev Ctries. 2014;8(08):961-972.
Mohebali M, Edrissian G, Nadim A et al. Application of direct agglutination test (DAT) for the diagnosis and seroepide-miological studies of visceral leishmaniasis in Iran. Iran J parasitol. 2006;1(1):15-25.
Mohebali M. Visceral leishmaniasis in Iran: review of the epidemiological and clinical features. Iran J parasitol. 2013;8(3):348.
Ndao M. Diagnosis of parasitic diseases: old and new approaches. Interdiscip Perspect Infect Dis. 2009;2009:1-16.
Mahmood T, Yang P-C. Western blot: technique, theory, and trouble shooting. N Am J Med Sci. 2012;4(9):429.
Berenguer J, Moreno S, Cercenado E et al. Visceral leishmaniasis in patients infected with human immunodeficiency virus(HIV). Ann Intern Med. 1989;111(2):129-132.
Salotra P, Raina A, Ramesh V. Western blot analysis of humoral immune response to Leishmania donovani antigens in patients with post-kala-azar dermal leishmaniasis. Trans R Soc Trop Med Hyg. 1999;93(1):98-101.
Dos Santos JI, Morgado MG, Galvão-Castro B. Human visceral leishmaniasis: analysis of the specificity of humoral immune response to polypeptides of Leishmania donovani chagasi. Am J Trop Med Hyg. 1987;37(2):263-270.
Persichetti MF, Solano-Gallego L, Vullo A et al. Diagnostic performance of ELISA, IFAT and Western blot for the detection of anti-Leishmania infantum antibodies in cats using a Bayesian analysis without a gold standard. Parasites & vectors. 2017;10(1):119.
Chappuis F, Sundar S, Hailu A et al. Visceral leishmaniasis: what are the needs for diagnosis, treatment and control?. Nature Rev Microbiol. 2007;5(11):873-882.
Mary C, Lamouroux D, Dunan S et al. Western blot analysis of antibodies to Leishmania infantum antigens: potential of the 14-kD and 16-kD antigens for diagnosis and epidemiologic purposes. Am J Trop Med Hyg. 1992;47(6):764-771.
Rolland-Burger L, Rolland X, Grieve C et al. Immunoblot analysis of the humoral immune response to Leishmania donovani infantum polypeptides in human visceral leishmaniasis. J Clin Microbiol. 1991;29(7):1429-1435.
Coelho VT, Oliveira JS, Valadares DG et al. Identification of proteins in promastigote and amastigote-like Leishmania using an immunoproteomic approach. PLoS Negl Trop Dis. 2012;6(1):e1430.
Rico AI, Gironès N, Fresno M et al. The heat shock proteins, Hsp70 and Hsp83, of Leishmania infantum are mitogens for mouse B cells. Cell Stress & Chaperones. 2002;7(4):339-346.
Shakarian AM, Joshi MB, Yamage M et al. Members of a unique histidine acid phosphatase family are conserved amongst a group of primitive eukaryotic human pathogens. Mol Cell Biochem. 2003;245(1):31-41.
Ashrafmansouri M, Sarkari B, Hatam G et al. Utility of Western Blot Analysis for the Diagnosis of Cutaneous Leishmaniasis. Iran J Parasitol. 2015;10(4):599.
Lopez M, Cherkasov A, Nandan D. Molecular architecture of leishmania EF-1α reveals a novel site that may modulate protein translation: A possible target for drug development. Biochem Biophys Res Commun. 2007;356(4):886-892.
Requena JM, Montalvo AM, Fraga J. Molecular chaperones of Leishmania: central players in many stress-related and-unrelated physiological processes. Bio Med Res Int. 2015;2015:1-21.
Kazemi-Rad E, Mohebali M, Khadem-Erfan MB et al. Overexpression of ubiquitin and amino acid permease genes in association with antimony resistance in Leishmania tropica field isolates. Korean J Parasitol. 2013;51(4):413-419.
Kumar P, Pai K, Tripathi K et al. Immunoblot analysis of the humoral immune response to Leishmania donovani polypeptides in cases of human visceral leishmaniasis: its usefulness in prognosis. Clin Diagn Lab Immunol. 2002;9(5):1119-23.
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