Publications by authors named "Diego H C Tavares"

2 Publications

  • Page 1 of 1

Gene design, optimization of protein expression and preliminary evaluation of a new chimeric protein for the serological diagnosis of both human and canine visceral leishmaniasis.

PLoS Negl Trop Dis 2020 07 27;14(7):e0008488. Epub 2020 Jul 27.

Instituto Aggeu Magalhães (IAM)-Fundação Oswaldo Cruz (Fiocruz), Recife, Pernambuco, Brazil.

Background: Visceral 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 Findings: Here, 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.

Conclusion: The 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.
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http://dx.doi.org/10.1371/journal.pntd.0008488DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7410341PMC
July 2020

A new recombinant F1 antigen as a cost and time-effective tool for plague diagnosis.

J Microbiol Methods 2020 05 27;172:105903. Epub 2020 Mar 27.

Departamento de Microbiologia - Instituto Aggeu Magalhães, Fiocruz, Recife, Pernambuco, Brazil. Electronic address:

The Yersinia pestis capsular antigen F1 is widely used in plague laboratory diagnosis. Here, we describe the production of an F1 recombinant protein within reduced time and biosafety requirements. Its evaluation in hemagglutination tests indicated that the recombinant F1 can replace the conventional F1 protein for plague diagnosis.
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http://dx.doi.org/10.1016/j.mimet.2020.105903DOI Listing
May 2020
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