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The increased presence of repetitive motifs in the KDDR-plus recombinant protein, a kinesin-derived antigen from Leishmania infantum, improves the diagnostic performance of serological tests for human and canine visceral leishmaniasis

PLoS Negl Trop Dis. 2021 Sep 17;15(9):e0009759. doi: 10.1371/journal.pntd.0009759. eCollection 2021 Sep.

Abstract

Visceral leishmaniasis (VL) is caused by protozoa belonging to the Leishmania donovani complex and is considered the most serious and fatal form among the different types of leishmaniasis, if not early diagnosed and treated. Among the measures of disease control stand out the management of infected dogs and the early diagnosis and appropriate treatment of human cases. Several antigens have been characterized for use in the VL diagnosis, among them are the recombinant kinesin-derived antigens from L. infantum, as rK39 and rKDDR. The main difference between these antigens is the size of the non-repetitive kinesin region and the number of repetitions of the 39 amino acid degenerate motif (6.5 and 8.5 repeats in rK39 and rKDDR, respectively). This repetitive region has a high antigenicity score. To evaluate the effect of increasing the number of repeats on diagnostic performance, we designed the rKDDR-plus antigen, containing 15.3 repeats of the 39 amino acid degenerate motif, besides the absence of the non-repetitive portion from L. infantum kinesin. Its performance was evaluated by enzyme-linked immunosorbent assay (ELISA) and rapid immunochromatographic test (ICT), and compared with the kinesin-derived antigens (rKDDR and rK39). In ELISA with human sera, all recombinant antigens had a sensitivity of 98%, whereas the specificity for rKDDR-plus, rKDDR and rK39 was 100%, 96% and 71%, respectively. When evaluated canine sera, the ELISA sensitivity was 97% for all antigens, and the specificity for rKDDR-plus, rKDDR and rK39 was 98%, 91% and 83%, respectively. Evaluation of the ICT/rKDDR-plus, using human sera, showed greater diagnostic sensitivity (90%) and specificity (100%), when compared to the IT LEISH (79% and 98%, respectively), which is based on the rK39 antigen. These results suggest that the increased presence of repetitive motifs in the rKDDR-plus protein improves the diagnostic performance of serological tests by increasing the specificity and accuracy of the diagnosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Protozoan / blood*
  • Dog Diseases
  • Dogs
  • Humans
  • Leishmania infantum*
  • Leishmaniasis, Visceral / diagnosis
  • Leishmaniasis, Visceral / parasitology
  • Leishmaniasis, Visceral / veterinary*
  • Protein Modification, Translational
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / genetics*
  • Recombinant Proteins
  • Sensitivity and Specificity
  • Serologic Tests / methods
  • Serologic Tests / veterinary*
  • Zoonoses

Substances

  • Antigens, Protozoan
  • Protozoan Proteins
  • Recombinant Proteins

Grants and funding

This study was supported by the following grants: RTF received financial support from Fundação de Amparo a Pesquisa do Estado de Minas Gerais/FAPEMIG, Brazil (http://www.fapemig.br) (Grant# APQ-04035-17 and APQ-02592-17); Conselho Nacional de Desenvolvimento Científico e Tecnológico/CNPq, Brazil (http://www.cnpq.br) (Grant# 303345/2018-7 and 421424/2018-4); and Pró-Reitoria de Pesquisa of Universidade Federal de Minas Gerais (https://www.ufmg.br/prpq). WFS has a PhD fellowship provided by FAPEMIG and Post-graduation Program in Infectious Diseases and Tropical Medicine/Universidade Federal de Minas Gerais. DCB, LLB and RTF are research fellows from CNPq/Brazil (Bolsa de Produtividade em Pesquisa). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.