Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Multicenter Study
. 2022 Jul 1:12:914477.
doi: 10.3389/fcimb.2022.914477. eCollection 2022.

Development of Immunological Assays Based on Leishmania donovani Antigen for Diagnosis of Canine Visceral Leishmaniasis and Their Multicenter Evaluation in Brazil and Italy

Sarfaraz Ahmad Ejazi  1 , Samiran Saha  1 , Anirban Bhattacharyya  1 , Sonali Das  1 , Nathália Lopes Fontoura Mateus  2 , Manoel Sebastião da Costa Lima  3 , Herintha Coeto Neitzke-Abreu  4 , Ivete Lopes de Mendonca  5 , Carlos Henrique Nery Costa  5 , Otoni Alves de Oliveira Melo  6 , Marcia Almeida de Melo  7 , Bartira Rossi-Bergmann  6 , Raffaele Corso  8 , Laura Manna  9 , Nahid Ali  1
Affiliations
Multicenter Study

Development of Immunological Assays Based on Leishmania donovani Antigen for Diagnosis of Canine Visceral Leishmaniasis and Their Multicenter Evaluation in Brazil and Italy

Sarfaraz Ahmad Ejazi et al. Front Cell Infect Microbiol. .

Abstract

Canine visceral leishmaniasis (CVL) due to Leishmania infantum infection is a zoonotic disease prevalent in the areas of South America and the Mediterranean. Infected dogs as reservoirs can contribute to disease transmission and can be a scourge to public health. Therefore, early diagnosis of infected dogs may play a pivotal role in circumscribing disease progression. Invasive tissue aspiration and insufficient serological methods impair a single assay for prompt CVL diagnosis. In the present study, we aimed to evaluate the potential of Leishmania donovani isolated membrane protein, LAg, for the diagnosis of CVL through immunological assays. Initially, enzyme-linked immunosorbent assay was done with Brazilian dog sera to evaluate the performance of LAg in diagnosing CVL and found sensitivity and specificity of 92.50% and 95%, respectively. The study further confirmed the diagnostic efficacy of LAg in a dipstick format. The dipstick test of canine sera from three centers in Brazil and one center in Italy collectively showed sensitivity values in the range of 53.33% to 100% in recognizing symptomatic dogs and specificity values between 75% and 100% to rule out healthy dogs. Moreover, a rapid immunochromatographic test was developed and optimized using LAg. This test was able to identify 94.73% of CVL of Brazilian origin with specificity of 97.29%. The current results highlight the reactive potential of the L. donovani antigen, LAg, for L. infantum CVL diagnosis and support our previous findings, which suggest the utility of LAg for the diagnosis of both L. donovani and L. infantum human VL in a variety of endemic regions. LAg as a diagnostic candidate may be employed to identify comprehensive CVL cases in epidemiological areas.

Keywords: Leishmania; canine; diagnosis; immunochromatographyic test (ICT); serology.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Indirect ELISA to detect antibodies in canine sera against leishmanial antigen LAg. (A) Brazilian CVL samples (n = 40) and healthy dogs sera (n = 40) were used in this study. Each dot represents an average value of a single sample. The significance between the groups was obtained as p < 0.0001 (***). (B) Cutoff was selected according to the receiver operator characteristic (ROC) curve obtained from GraphPad Prism 5.
Figure 2
Figure 2
Representative result of dipstick test with positive and negative CVL sera.
Figure 3
Figure 3
Representative result of ICT with positive and negative CVL sera.
See this image and copyright information in PMC

References

    1. Abbehusen M., Almeida V., Solcà M., Pereira L., Costa D. J., Gil-Santana L., et al. . (2017). Clinical and Immunopathological Findings During Long Term Follow-Up in Leishmania Infantum Experimentally Infected Dogs. Sci. Rep. 7 (1), 15914. doi: 10.1038/s41598-017-15651-8 - DOI - PMC - PubMed
    1. Aparecida de Carvalho C., Mitsuyoshi Hiramoto R., Regina Meireles L., Franco de Andrade Júnior H. (2021). Serum Antibodies Blocked by Glycan Antigens in Canine Visceral Leishmaniasis Serology are Mostly IgA Immune Complexes. Parasitology 148 (12), 1509–1515. doi: 10.1017/S0031182021001189 - DOI - PMC - PubMed
    1. Barbosa de Castro R., Badaró de Moraes J. V., Costa Bressan G., de Souza Vasconcellos R., Silva-Júnior A., Lopes Rangel Fietto J. (2022). Antigens and Their Diagnostic Performance for Canine Visceral Leishmaniasis: A Critical Review. Vet. Parasitol . 301 109638. doi: 10.1016/j.vetpar.2021.109638 - DOI - PubMed
    1. Contreras M. E. M. (2018). Serological Diagnosis of Canine Visceral Leishmaniasis. Am. J. Clin. Microbiol. Antimicrob. 1 (3).
    1. Cota G. F., de Sousa M. R., Demarqui F. N., Rabello A. (2012). The Diagnostic Accuracy of Serologic and Molecular Methods for Detecting Visceral Leishmaniasis in HIV Infected Patients: Meta-Analysis. PLoS Negl. Trop. Dis. 6 (5), e1665. doi: 10.1371/journal.pntd.0001665 - DOI - PMC - PubMed

Publication types

LinkOut - more resources