Immunodiagnosis of canine visceral leishmaniasis using mimotope peptides selected from phage displayed combinatorial libraries.

Christina Monerat Toledo-Machado,Carlos Chávez-Olórtegui,Ricardo Toshio Fujiwara,Claudia Martins Carneiro,Lilian Lacerda Bueno,Christophe Nguyen,Rubens Antonio Carneiro,Ricardo Andrez Machado De Avila,Daniel Menezes-Souza,Claude Granier

doi:10.1155/2015/401509

Abstract

ELISA and RIFI are currently used for serodiagnosis of canine visceral leishmaniasis (CVL). The accuracy of these tests is controversial in endemic areas where canine infections by Trypanosoma cruzi may occur. We evaluated the usefulness of synthetic peptides that were selected through phage display technique in the serodiagnosis of CVL. Peptides were chosen based on their ability to bind to IgGs purified from infected dogs pooled sera. We selected three phage clones that reacted only with those IgGs. Peptides were synthesized, polymerized with glutaraldehyde, and used as antigens in ELISA assays. Each individual peptide or a mix of them was reactive with infected dogs serum. The assay was highly sensitive and specific when compared to soluble Leishmania antigen that showed cross-reactivity with anti-T. cruzi IgGs. Our results demonstrate that phage display technique is useful for selection of peptides that may represent valuable synthetic antigens for an improved serodiagnosis of CVL.

Highlights

Canine visceral leishmaniasis (CVL) caused by Leishmania (Leishmania) infantum chagasi is a widespread zoonotic disease of both the Old and the New World [1] leading to a considerable number of deaths
We evaluated the usefulness of synthetic peptides that were selected through phage display technique in the serodiagnosis of canine visceral leishmaniasis (CVL)
In order to estimate the diagnostic accuracy, we found three peptides that could successfully be used as antigens in ELISA assays for a specific immunodiagnosis of canine visceral leishmaniasis without cross reaction with circulating antibodies of T. cruzi experimentally infected dogs

Summary

Introduction

Canine visceral leishmaniasis (CVL) caused by Leishmania (Leishmania) infantum chagasi is a widespread zoonotic disease of both the Old and the New World [1] leading to a considerable number of deaths. The Brazilian Ministry of Health recommends the use of an immunoenzymatic assay (ELISA) and an indirect immunofluorescence antibody test (IFAT) for the diagnosis of canine visceral leishmaniasis (CVL), employed as criteria for the culling of seropositive dogs in surveillance and control programs for visceral leishmaniasis (VL) [10]. Both the accuracy of these tests and the process of dog culling promote a controversial impact of the leishmaniasis infection [11, 12]. The development of an effective diagnosis test can be critical for the control and the possible eradication of VL; BioMed Research International more sensitive and specific tests may be especially helpful to achieve this goal [13]

Methods

Results

Conclusion