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Potential application of rLc36 protein for diagnosis of canine visceral leishmaniasis.

ABSTRACT: Visceral leishmaniasis (VL) is fatal if left untreated. Infected dogs are important reservoirs of the disease, and thus specific identification of infected animals is very important. Several diagnostic tests have been developed for canine VL (CVL); however, these tests show varied specificity and sensitivity. The present study describes the recombinant protein rLc36, expressed by Leishmania infantum, as potential antigen for more sensitive and specific diagnosis of CVL based on an immunoenzymatic assay. The concentration of 1.0 ?g/mL of rLc36 enabled differentiation of positive and negative sera and showed a sensitivity of 85% and specificity of 71% (with 95% confidence), with an accuracy of 76%.

SUBMITTER: Nogueira CT

PROVIDER: S-EPMC5804312 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature

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Potential application of rLc36 protein for diagnosis of canine visceral leishmaniasis.

Nogueira Camila Tita CT Cistia Mayara Lúcia Del MLD Urbaczek Ana Carolina AC Jusi Márcia Mg MM Velásquez Angela Maria Arenas AMA Machado Rosângela Zacarias RZ Ferreira Henrique H Henrique-Silva Flávio F Langoni Hélio H Costa Paulo Inácio da PID Graminha Márcia As MA

Memorias do Instituto Oswaldo Cruz 20180301 3

Visceral leishmaniasis (VL) is fatal if left untreated. Infected dogs are important reservoirs of the disease, and thus specific identification of infected animals is very important. Several diagnostic tests have been developed for canine VL (CVL); however, these tests show varied specificity and sensitivity. The present study describes the recombinant protein rLc36, expressed by Leishmania infantum, as potential antigen for more sensitive and specific diagnosis of CVL based on an immunoenzymati ...[more]

PMID: 29412359

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Project description:BackgroundVisceral leishmaniasis (VL) is a major neglected disease, potentially fatal, whose control is still impaired by inefficient and/or expensive treatment and diagnostic methods. The most promising approach for VL diagnosis uses serological assays with recombinant proteins, since they are more efficient and easier to perform. Tests developed for the human form of the disease, however, have not been shown to be efficient for its diagnosis in the canine host, the major reservoir for the American VL.Methodology/principal findingsHere, we describe a systematic approach aimed at the production of a new chimeric protein potentially able to be used for both human and canine VL diagnosis and based both on in silico gene design and experimental data. Starting from the previous identification of Leishmania infantum recombinant antigens efficient for the diagnosis of either human or canine VL, three of the best performing antigens were selected (Lci2, Lci3 and Lci12). After a preliminary evaluation validating the chimeric approach, DNA fragments encoding predicted antigenic regions from each protein, enriched with repeats, were joined in various combinations to generate a total of seventeen chimeric genes optimized for prokaryotic expression. These were assessed for optimal expression and purification yield, with four chimeric proteins being efficiently produced. Their diagnostic potential was then evaluated through ELISA assays with sera from VL afflicted humans and dogs. After two rounds of gene design, the results showed high levels of sensitivity for the best chimeric protein, named Q5, in humans (82%) and dogs (100%) with 100% specificity in comparison with healthy controls. A single non-specific reaction was seen with serum from individuals with tegumentary leishmaniasis.ConclusionThe newly described chimeric protein is potentially useful for the detection of both humans and dogs afflicted with VL, with its use in rapid tests necessary for validation as a new diagnostic tool.

Dataset Information

Potential application of rLc36 protein for diagnosis of canine visceral leishmaniasis.

Publications

Potential application of rLc36 protein for diagnosis of canine visceral leishmaniasis.

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