Veterinary Sciences (Oct 2023)

Diagnostic Potential for the Detection of Canine Visceral Leishmaniasis of an ELISA Assay Based on the Q5 Recombinant Protein: A Large-Scale and Comparative Evaluation Using Canine Sera with a Positive Diagnosis from the Dual-Path-Platform (DPP) Test

  • Larissa Ferreira de Araújo Paz,
  • Adalúcia da Silva,
  • Hemilly Rayanne Ferreira da Silva,
  • Milena Paiva Cavalcanti,
  • Valeria Marçal Felix de Lima,
  • Maria Rosário Oliveira da Cunha Beltrão,
  • Maria Beatriz Araújo Silva,
  • Osvaldo Pompílio de Melo Neto,
  • Zulma Maria Medeiros,
  • Wagner José Tenório dos Santos

DOI
https://doi.org/10.3390/vetsci10100608
Journal volume & issue
Vol. 10, no. 10
p. 608

Abstract

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Dogs are considered the major domestic reservoir for human visceral leishmaniasis, a serious disease caused by the Leishmania infantum parasite. Diagnosis of canine visceral leishmaniasis (CVL) is critical for disease control, with several methods currently available. Among the serological tests, the DPP rapid test and the EIE-LVC, more commonly used in Brazil, are associated with variable sensitivity and specificity. Research with novel recombinant proteins such as the ELISA with the recombinant chimeric protein Q5 may therefore improve the CVL diagnosis. This study aimed to evaluate the true diagnostic potential of Q5 in an ELISA assay using a large number of CVL-suspected sera (406) with a previous positive diagnosis based on the rapid DPP test. Sera from the DPP-positive dogs, also assessed with the EIE-LVC test, were compared with sera from healthy dogs (n = 46) and used for ELISA tests using the recombinant Q5. The resulting data as well as the correlation with the clinical signs and the environmental characteristics of the animals were analyzed using Medal and GraphPad Prism 8.0. Overall, similar levels of lower sensitivity (67–68%) were seen for both the commercial EIE-LVC test and the Q5 ELISA when all assessed sera were considered, but a much greater sensitivity (92%) was seen for those samples from symptomatic dogs only. In contrast, many negative results were observed for the DPP-positive sera from asymptomatic dogs or those with no clinical information available. A selection of those sera were tested yet again in new ELISA assays using a second batch of the recombinant Q5, purified under milder denaturing conditions, as well as using another recombinant protein (Lci13). The results reveal a higher-than-expected incidence of likely false-positive results for DPP, reinforcing the need for other recombinant proteins, such as the chimeric Q5, to be investigated as possible alternatives to the currently used CVL diagnostic methods.

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