Multi-antigen print immunoassay (MAPIA)-based evaluation of novel recombinant Leishmania infantum antigens for the serodiagnosis of canine visceral leishmaniasis.

Isaac Queiroz De Oliveira,Edmilson Domingos Da Silva,Deborah Bittencourt Mothé Fraga,Alexandre Barbosa Reis,Gabriel Grimaldi,Rodrigo Araujo Silva,Michel Vergne Sucupira,Patrícia Sampaio Tavares Veras

doi:10.1186/s13071-015-0651-6

Abstract

BackgroundDomestic dogs are the principal reservoir hosts of Leishmania infantum in regions where visceral leishmaniasis (VL) is endemic. Although serologic methods are frequently used for the screening of infected dogs, antibody-based tests require further assessment, due to lack of sensitivity and specificity. In this study, we employed a multi-antigen printing immunoassay (MAPIA) to compare the antibody responses to novel recombinant proteins of L. infantum with the potential for the detection of canine VL.FindingsMAPIA strips were prepared employing 12 recombinant proteins. Antibody reactivity to these antigens was compared using a panel of sera collected from clinically asymptomatic (n = 16) and symptomatic (n = 41) culture-positive animals. Our findings showed that the canine immune response to antigen differs between dogs and depends on infection status. Using this screening assay, when five out of the 12 antigens were combined, an overall 81% detection rate of L. infantum-infected dogs was achieved.ConclusionsWe conclude that MAPIA is an effective screening tool to rapidly select multiple antigens of diagnostic utility to be used in a more sensitive point of care diagnostic test such as the Dual-Path Platform (DPP) multiplex test for the rapid detection of infected dogs.

Highlights

Domestic dogs are the principal reservoir hosts of Leishmania infantum in regions where visceral leishmaniasis (VL) is endemic
We conclude that multi-antigen printing immunoassay (MAPIA) is an effective screening tool to rapidly select multiple antigens of diagnostic utility to be used in a more sensitive point of care diagnostic test such as the Dual-Path Platform (DPP)
Parasite-specific antibody tests, such as the immunofluorescent-antibody test [IFAT], direct agglutination test [DAT], enzyme-linked immunosorbent assay [ELISA], widely used to diagnose infection, employ crude antigens derived from wholeparasite extracts and lack the appropriate sensitivity and specificity required for accurate serodiagnosis [5,6,7,8,9,10,11]

Summary

Introduction

Domestic dogs are the principal reservoir hosts of Leishmania infantum in regions where visceral leishmaniasis (VL) is endemic. To test for crossreactivity of the 12 recombinant antigens with other pathogens, we screened sera from dogs infected with Leishmania braziliensis (n = 10), Trypanosoma cruzi (n = 10), Babesia spp. The antibody reactivity was assessed using a panel of 57 sera from symptomatic (n = 41) and asymptomatic (n = 16) culture-positive dogs.

Results

Conclusion