Article
High Performance of ELISA test using recombinant rLiNTPDase2 from Leishmania infantum: a Phase II diagnosis of Canine Visceral Leishmaniasis
Registro en:
SOUZA, Anna Cláudia Alves de et al. High Performance of ELISA test using recombinant rLiNTPDase2 from Leishmania infantum: a Phase II diagnosis of Canine Visceral Leishmaniasis. Acta Tropica, v. 209, 105535, p. 1-10, 2020.
0001-706X
10.1016/j.actatropica.2020.105535
1873-6254
Autor
Souza, Anna Cláudia Alves de
Castro, Raissa Barbosa de
Santos, Yaro Luciolo dos
Pavione, Nancy da Rocha Torres
Agripino, Joice de Melo
Bahia, Maria Terezinha
Coelho, George Luiz Lins Machado
Souza, Ronny Francisco de
Oliveira, Leandro Licursi de
Souza, Celeste da Silva Freitas de
Bressan, Gustavo Costa
Vasconcellos, Raphael de Souza
Almeida, Márcia Rogéria de
Fietto, Juliana Lopes Rangel
Resumen
Canine visceral leishmaniasis (CVL) has been the theme of several studies given the importance of dog as natural
reservoir of the pathogen Leishmania infantum in endemic regions and its role on dissemination of CVL and
human visceral Lesihmaniasis (VL). The current immunodiagnosis of CVL has limitations concerning accuracy,
specificity and sensitivity. Therefore, improvements are required. rLiNTPDase2 has been previously highlighted
as a new recombinant antigen from L. infantum to the CVL diagnosis by ELISA assay (rLiNTPDase2-ELISA). In this
study, we aimed to evaluate rLiNTPDase2-ELISA in a Phase II study with 651 dog sera samples, also comparing it
with methodologies previously established and used in epidemiology surveillance in Brazil, an endemic country
of CVL and VL. The rLiNTPDase2-ELISA using standard control sera showed high capability to distinguish between positive and negative sera, sensitivity of 92.6% and specificity of 88.5%. The test was reproductive and
the kappa statistics judgement “substantial agreement”. rLiNTPDase2-ELISA does not show cross-reactivity with
ehrlichiosis-reagent sera. However, we verified 15.3% of cross-reactivity with Chagas disease-reagent sera. The
performance of rLiNTPDase2-ELISA was evaluated using sera samples from vaccinated dogs (Leish-Tec®). The
results showed high agreement with parasitological and PCR results (sensitivity of 100.0% and specificity of
91.7%). Furthermore, we compared the performance of rLiNTPDase2-ELISA in CVL-reagent sera samples from
endemic areas, which were previously diagnosed using other tests for CVL: immunofluorescent (IFI-LVC-BioManguinhos), IFI-LVC-Bio-Manguinhos coupled to ELISA (EIE-LVC-Bio-Manguinhos) and the Rapid Dual Path
Platform® (TR-DPP®-Bio-Manguinhos) coupled to EIE-LVC-Bio-Manguinhos. rLiNTPDase2-ELISA showed high
level of concordance with IFI-LVC-Bio-Manguinhos (88.6%) and with IFI-LVC-Bio-Manguinhos coupled to EIELVC-Bio-Manguinhos (82.9%) but not with TR-DPP® -Bio-Manguinhos coupled to EIE-LVC-Bio-Manguinhos
(33.3%), which casts doubts on the effectiveness of this latest test. In addition, the rLiNTPDase2 antigen adsorbed in 96-well plate was stable enough to be used at least for three months. Taken together, our data confirmed, by Phase II study using hundreds samples, the good potential of rLiNTPDase2-ELISA to be used in the
field as a new diagnostic assay for CVL. 2025-01-01