Chemiluminescent ELISA with multi-epitope proteins to improve the diagnosis of canine visceral leishmaniasis

Fonseca, Thaisa Helena Silva; Faria, Angélica Rosa; Leite, Higor; Silveira, Júlia A.G.; Carneiro, Cláudia Martins; Andrade, Hélida Monteiro de

doi:10.1016/j.tvjl.2019.105387

Cited by 15 publications

(8 citation statements)

References 36 publications

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“…In comparison with a similar study in Brazil, crude antigen has demonstrated 75% sensitivity and 73.3% specificity in chemiluminescent ELISA. However, with a multi-epitope protein, PQ10, the sensitivity and specificity have been reported to be 93% and 80%, respectively (Fonseca et al, 2019). Therefore, in the preliminary investigation, LAg-ELISA showed acceptable sensitivity and specificity in identifying canine VL.…”

Section: Discussionmentioning

confidence: 86%

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

Ejazi

Saha

Bhattacharyya

et al. 2022

Front. Cell. Infect. Microbiol.

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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.

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Section: Discussionmentioning

confidence: 86%

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

Ejazi

Saha

Bhattacharyya

et al. 2022

Front. Cell. Infect. Microbiol.

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“…In this study, the dog samples were submitted to different tests (IFA and DPP ® ) to confirm their Leishmania infection. Recombinant proteins produce the best ELISA results for diagnosing canine visceral leishmaniosis when compared with crude extracted antigens (FARIA et al, 2015;FONSECA et al, 2019). However, the ease and availability of obtaining crude antigens allow for the standardization of ELISA and its practical application in regions with fewer resources.…”

Section: Discussionmentioning

confidence: 99%

In-house serological ELISA as a leishmaniosis diagnostic test: development and applications in canines from the western border of Brazil

et al. 2023

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Leishmaniosis is a great public health problem affecting both humans and animals. The disease is caused by the protozoan Leishmania spp., which has a complex cycle involving a phlebotomine vector. The ELISA test (Enzyme-Linked Immunosorbent Assay) along with a chromatographic immunoassay was defined by the Brazil Health Ministry as the confirmatory screening protocol in 2011. Uruguaiana city is 630 km away from Porto Alegre, which makes it difficult to send samples and diagnose leishmaniasis, as well as receive quick results. In view of this, the present study evaluated an in-house indirect ELISA method compared to indirect immunofluorescence assay (IFA) and dual-path platform chromatographic immunoassay (DPP-BioManguinhos®) for the detection of an immune response to Leishmania spp. in canine species. The serological evaluation included 48 canines from the western border of Brazil (Uruguaiana and Barra do Quaraí city). Among the 48 canine samples tested, 18 were positive when using the ELISA technique, 19 were positive with IFA, and 17 were positive with rapid test DPP®. The ELISA technique showed a sensitivity/specificity of 83.3%/86.7% when compared to IFA and 100%/96.8% compared to DPP®. The present study showed a prevalence of 37.5%, demonstrating that the infection circulates in the studied population. It can be concluded that the ELISA technique was valuable for use in field conditions when performing screening tests in endemic areas.

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“…infantum -positive serum samples/30 healthy dog serum samples/32 serum samples from non- L . infantum diseases PQ10 Sensitivity: 93.1% Specificity: 80% PQ20 Sensitivity: 93.1% Specificity: 96.6% [ 79 ]/Brazil Canine visceral leishmaniasis/ Leishmania infantum PQ10 E. coli BL21(DE3) cells ELISA/ Direct agglutination 50 L . infantum -positive serum samples/50 healthy dog serum samples/30 serum samples from non- L .…”

Section: Rmps Applied In Disease Diagnosismentioning

confidence: 99%

Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis

Gonçalves,

Ribeiro,

Resende

et al. 2024

Microb Cell Fact

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Recombinant multiepitope proteins (RMPs) are a promising alternative for application in diagnostic tests and, given their wide application in the most diverse diseases, this review article aims to survey the use of these antigens for diagnosis, as well as discuss the main points surrounding these antigens. RMPs usually consisting of linear, immunodominant, and phylogenetically conserved epitopes, has been applied in the experimental diagnosis of various human and animal diseases, such as leishmaniasis, brucellosis, cysticercosis, Chagas disease, hepatitis, leptospirosis, leprosy, filariasis, schistosomiasis, dengue, and COVID-19. The synthetic genes for these epitopes are joined to code a single RMP, either with spacers or fused, with different biochemical properties. The epitopes’ high density within the RMPs contributes to a high degree of sensitivity and specificity. The RMPs can also sidestep the need for multiple peptide synthesis or multiple recombinant proteins, reducing costs and enhancing the standardization conditions for immunoassays. Methods such as bioinformatics and circular dichroism have been widely applied in the development of new RMPs, helping to guide their construction and better understand their structure. Several RMPs have been expressed, mainly using the Escherichia coli expression system, highlighting the importance of these cells in the biotechnological field. In fact, technological advances in this area, offering a wide range of different strains to be used, make these cells the most widely used expression platform. RMPs have been experimentally used to diagnose a broad range of illnesses in the laboratory, suggesting they could also be useful for accurate diagnoses commercially. On this point, the RMP method offers a tempting substitute for the production of promising antigens used to assemble commercial diagnostic kits.

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Chemiluminescent ELISA with multi-epitope proteins to improve the diagnosis of canine visceral leishmaniasis

Cited by 15 publications

References 36 publications

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

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

In-house serological ELISA as a leishmaniosis diagnostic test: development and applications in canines from the western border of Brazil

Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis

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