Utility of Western Blot Analysis for the Diagnosis of Cutaneous Leishmaniasis
AbstractBackground: Cutaneous leishmaniasis (CL) is a parasitic disease with a relatively wide distribution in different areas of the world, including Iran. The parasite is mainly diagnosed microscopically, but serological approaches might be useful for diagnosis as well. This study aimed to assess the efficacy of an immunoblotting system for serodiagnosis of cutaneous leishmaniasis in Iran.Methods: Sixty-one sera samples from parasitologically confirmed CL patients and 50 sera samples from healthy controls along with 50 sera sample from non-CL patients were collected. Native strain of Leishmania major was cultured in Schneider medium and soluble Leishmania antigens were prepared from amastigotes-like parasites. All of sera samples were evaluated by an immunoblotting system.Results: Components of 14 to 135 kDa were detectable by the sera of CL patients. From 61 sera of CL patients, 59 cases (96.7%) detected a 63 kDa subunit and 51 cases (83.6%) recognized a 32-35 kDa component. Among all subunits, the 63 kDa band showed the highest sensitivity (96.7%) and a 75 kDa band had the highest (98%) specificity.Conclusion: Immunoblotting has a satisfactory performance in diagnosis of CL and this test can be used, as an aid, for proper diagnosis of CL.
Alvar J, Velez ID, Bern C, Herrero M, Desjeux P, Cano J, et al. Leishmaniasis worldwide and global estimates of its incidence. PLoS One. 2012; 7(5):e35671.
Motazedian H, Noamanpoor B, Ardehali S. Characterization of Leishmania parasites isolated from provinces of the Islamic Republic of Iran. East Mediterr Health J. 2002; 8(2-3):338-344.
Hajjaran H, Mohebali M, Mamishi S, Vasigheh F, Oshaghi MA, Naddaf SR, et al. Molecular identification and polymorphism determination of cutaneous and visceral leishmaniasis agents
isolated from human and animal hosts in Iran. Biomed Res Int. 2013; 2013:789326.
Mohebali M. Visceral leishmaniasis in Iran: Review of the epidemiological and clinical fea-tures. Iran J Parasitol. 2013; 8(3):348-358.
Sarkari B, Hatam G, Ghatee M. Epidemiologi-cal features of visceral leishmaniasis in Fars province, southern Iran. Iran J Public Health. 2012; 41(4):94-99.
Souza AP, Soto M, Costa JM, Boaventura VS, de Oliveira CI, Cristal JR, et al. Towards a more precise serological diagnosis of human tegumentary leishmaniasis using Leishmania re-combinant proteins. PLoS One. 2013; 8(6):e66110.
Singh S. New developments in diagnosis of leishmaniasis. Indian J Med Res. 2006; 123(3):311-30.
Reithinger R, Dujardin JC. Molecular diagnosis of leishmaniasis: current status and future ap-plications. J Clin Microbiol. 2007; 45(1):21-25.
Goto H, Lindoso JA. Current diagnosis and treatment of cutaneous and mucocutaneous leishmaniasis. Expert Rev Anti Infect Ther. 2010; 8(4):419-433.
Ameen M. Cutaneous leishmaniasis: advances in disease pathogenesis, diagnostics and thera-peutics. Clin Exp Dermatol. 2010; 35(7):699-705.
Srividya G, Kulshrestha A, Singh R, Salotra P. Diagnosis of visceral leishmaniasis: develop-ments over the last decade. Parasitol Res. 2012; 110(3):1065-1078.
Mikaeili F, Fakhar M, Sarkari B, Motazedian MH, Hatam G. Comparison of serological methods (ELISA, DAT and IFA) for diagnosis of visceral leishmaniasis utilizing an endemic strain. Iran J Immunol 2007; 4(2):116-121.
Sarkari B, Hatam GR, Mikaeili F, Sadeghi H, Ebrahimi S. A comparative study of antigen and antibody detection in visceral leishmaniasis using serum and urine-based ELISA. Trop Bi-omed. 2008; 25(2):96-99.
Ghatei MA, Hatam GR, Hossini MH, Sarkari B. Performance of latex agglutination test (KA-tex) in diagnosis of visceral leishmaniasis in Iran. Iran J Immunol. 2009; 6(4):202-207.
Castellano LR, Gave TC, Lira MA, Dessein H, Dessein A, Correia D, et al. Evaluation of elec-tro-eluted antigens in the serological diagnosis of cutaneous leishmaniasis. Ann Trop Med Parasitol. 2010; 104(4):347-350.
Szargiki R, Castro EA, Luz E, Kowalthuk W, Machado AM, Thomaz-Soccol V. Comparison of serological and parasitological methods for cutaneous leishmaniasis diagnosis in the state of Parana, Brazil. Braz J Infect Dis. 2009; 13(1):47-52.
Al-Nahhas SA. Serodiagnosis of cutaneous leishmaniasis in the Syrian Arab Republic. Sau-di Med J. 2009; 30(3):382-386.
Barroso-Freitas AP, Passos SR, Mouta-Confort E, Madeira MF, Schubach AO, Santos GP et al. Accuracy of an ELISA and indirect immuno-fluorescence for the laboratory diagnosis of American tegumentary leishmaniasis. Trans R Soc Trop Med Hyg. 2009; 103(4):383-389.
Zeyrek FY, Korkmaz M, Ozbel Y. Serodiagno-sis of anthroponotic cutaneous leishmaniasis (ACL) caused by Leishmania tropica in Sanliurfa Province, Turkey, where ACL is highly en-demic. Clin Vaccine Immunol. 2007; 14(11):1409-1415.
Ribeiro FC, de OS, Mouta-Confort E, Schu-bach TM, de Fatima MM, Marzochi MC. Use of ELISA employing Leishmania (Viannia) braziliensis and Leishmania (Leishmania) chagasi an-tigens for the detection of IgG and IgG1 and IgG2 subclasses in the diagnosis of American tegumentary leishmaniasis in dogs. Vet Parasi-tol. 2007; 148(3-4):200-206.
Rocha RD, Gontijo CM, Eloi-Santos SM, Teixeira-Carvalho A, Correa-Oliveira R, Ferrari TC, et al. Clinical value of anti-live Leishmania (Viannia) braziliensis immunoglobulin G sub-classes, detected by flow cytometry, for diag-nosing active localized cutaneous leishmaniasis. Trop Med Int Health. 2006; 11(2):156-166.
Habibi P, Sadjjadi SM, Owji M, Moattari A, Hatam GR, Sarkari B. Characterization of in vitro cultivated amastigote like of Leishmania major: a substitution for in vivo studies. Iran J Parasitol. 2008; 3(1):6-15.
Jensen AT, Gaafar A, Ismail A, Christensen CB, Kemp M, Hassan AM et al. Serodiagnosis of cutaneous leishmaniasis: assessment of an enzyme-linked immunosorbent assay using a peptide sequence from gene B protein. Am J Trop Med Hyg, 1996; 55(5):490-495.
Romero LI, Paz HM, Ortega-Barria E, Bayard V, Hochberg LP, Collins KM et al. Evaluation of serological assays based on a novel excreted antigen preparation for the diagnosis of cutane-ous leishmaniasis in Panama. J Microbiol Methods. 2004; 57(3):391-397.
Sarkari B, Ashrafmansouri M, Hatam G, Habi-bi P, Abdolahi Khabisi S. Performance of an ELISA and indirect immunofluorescence assay in serological diagnosis of zoonotic cutaneous leishmaniasis in Iran. Interdiscip Perspect In-fect Dis. 2014; 2014:505134.
Lieke T, Nylén S, Eidsmo L, McMaster WR, Mohammadi AM, Khamesipour A, Berg L, Akuffo H. Leishmania surface protein gp63 binds directly to human natural killer cells and inhibits proliferation. Clin Exp Immunol. 2008; 153(2):221-30.
Pomares C, Despierres L, del GP, Delaunay P, Michel G, Ferrua B et al. Western blot analysis as an aid for the diagnosis of cutaneous leish-maniasis due to Leishmania major. Trans R Soc Trop Med Hyg. 2012; 106(7):452-454.
Masuda A, do Nascimento SF, Guerra CS, Paranhos GS, Ferreira AW. Analysis of the specificity of human antibodies to antigens of Leishmania braziliensis braziliensis. Rev Inst Med Trop Sao Paulo. 1989; 31(4):228-234.