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Evaluation of Leishmania Species Reactivity in Human Serologic Diagnosis of Leishmaniasis

Ricardo Silvestre, Nuno Santarém, Lúcia Teixeira, Joana Cunha, Henk Schallig, AND Anabela Cordeiro-da-Silva^*

The sensitivities and specificities of IgG-ELISA and IgG flow cytometry based techniques using different Leishmania species were determined using sera collected from 40 cutaneous or visceral leishmaniasis patients. The flow cytometry technique, using promastigote parasite forms, performed better than total soluble extract IgG-ELISA. At the species level, the use of Leishmania amazonensis and Leishmania major as antigens in enzyme linked immunosorbent assay (ELISA) decreased the overall sensitivity. To assess the specificity of these tests, sera from malaria, toxoplasmosis, amoebiasis, schistosomiasis, and leprosy patients were used. We also included sera from Leishmania non-infected endemic individuals. The cutaneous species displayed a decreased specificity in both assays. Although more sensitive, flow cytometry using promastigote parasite forms generally presented lower levels of specificity when compared with total extract of IgG-ELISA. Overall, the results of the study show the potential of IgG flow cytometry for the diagnosis of leishmaniasis. Although highly sensitive, a refinement of the flow cytometry method should be performed to improve the overall specificity.

Received September 3, 2008. Accepted for publication April 20, 2009.

Financial support: R. Silvestre and N. Santarém were supported by fellowships from FCT, POCI 2010, and co-funded by FEDER in the project SFRH/BPD/41476/2007 and POCI/SAU-FCT/59837/2004, respectively. The work was also supported by FCT, POCI 2010, and co-funded by FEDER in the project PTD/SAU-FCF/67351/2006 and PTDC/CVT/65047/2006

Disclosure: This study was performed as part of a reviewed and approved protocol by the Medical Ethical Committee of the Academic Medical Center in Amsterdam (MEC 03/228) in 2003.

^* Address correspondence to Anabela Cordeiro-da-Silva, Parasite Disease Group, Biology of Infection and Immunology, Instituto de Biologia Molecular e Celular da Universidade do Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal. E-mail: cordeiro{at}ibmc.up.pt

These authors contributed equally to this work.

Authors’ addresses: Ricardo Silvestre, Nuno Santarém, Lúcia Teixeira, Joana Cunha, and Anabela Cordeiro-da-Silva, Parasite Disease Group, Biology of Infection and Immunology, IBMC—Instituto de Biologia Molecular e Celular, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal, Tel: +351-226-074-900, Fax: +351-226-099-157, and Departamento de Bioquímica, Faculdade de Farmácia, Universidade do Porto, Portugal, Rua Aníbal Cunha, 164, 4050-047 Porto, Tel: 00351-222-078-900, Fax: 00351-222-003-977, E-mails: rleal{at}ibmc.up.pt , nsantarem{at}portugalmail.pt , jcunha{at}ibmc.up.pt , luciat{at}ibmc.up.pt , and cordeiro{at}ibmc.up.pt . Henk Schallig, KIT (Koninklijk Instituut voor de Tropen/Royal Tropical Institute), KIT Biomedical Research, Meibergdreef 39, 1105 AZ Amsterdam, The Netherlands, Tel: 0031(0)20-5665447, Fax: 0031(0)20-6971841, E-mail: h.schallig{at}kit.nl .

Reprint requests: Anabela Cordeiro-da-Silva, Parasite Disease Group, Biology of Infection and Immunology, Instituto de Biologia Molecular e Celular da Universidade do Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal, E-mail: cordeiro{at}ibmc.up.pt.