Potential application of small myristoylated protein-3 evaluated as recombinant antigen and a synthetic peptide containing its linear B-cell epitope for the serodiagnosis of canine visceral and human tegumentary leishmaniasis

Salles, Beatriz C.S.; Dias, Daniel Lucas Santos; Steiner, Bethina T.; Lage, Daniela P.; Ramos, Fernanda F.; Ribeiro, Patrícia A.F.; Santos, Thaís T.O.; Lima, Mariana P.; Costa, Lourena E.; Chaves, Ana Cristina; Chávez-Fumagalli, Miguel Á.; Fujiwaraa, Ricardo T.; Buenoa, Lílian L.; Caligiorne, Rachel Basques; Magalhães-Soares, Danielle F. de; Silveira, Júlia A.G.; Machado-de-Ávila, Ricardo Andrez; Gonçalves, Denise Utsch; Coelho, Eduardo Antônio Ferraz

doi:10.1016/j.imbio.2018.09.003

Cited by 16 publications

(13 citation statements)

References 58 publications

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“…Immunological methods have been also employed; however, problems related with the sensitivity and/or specificity of the antigens used in the tests are registered [11,12]. Thus, it is necessary to search for new candidates that will serve to design diagnostic systems with higher degree of sensitivity and specificity than current methods, such as the refinement of use of new antigens, such as recombinant proteins [13,14,15], synthetic peptides [16,17], polypeptide-based chimera [18,19] and phage clones [8,20].…”

Section: Introductionmentioning

confidence: 99%

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

Costa

Alves

Carneiro

et al. 2019

IJMS

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Two Leishmania infantum mimotopes (B10 and C01) identified by phage display showed to be antigenic and immunogenic for visceral (VL) and tegumentary (TL) leishmaniasis; however, their biological targets in the parasites have not been identified. The aim of the present study was to investigate the native antigens expressing both mimotopes, and to use them in distinct immunological assays. For this, a subtractive phage display technology was used, where a combinatorial library of single-chain variable fragments (scFv) was employed and the most reactive monoclonal antibodies for each target were captured, being the target antigens identified by mass spectrometry. Results in immunoblotting and immunoprecipitation assays showed that both monoclonal scFvs antibodies identified the β-tubulin protein as the target antigen in L. infantum. To validate these findings, the recombinant protein was cloned, purified and tested for the serodiagnosis of human leishmaniasis, and its immunogenicity was evaluated in PBMC derived from healthy subjects and treated or untreated VL patients. Results showed high diagnostic efficacy, as well as the development of a specific Th1 immune response in the cell cultures, since higher IFN-γ and lower IL-10 production was found.

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Section: Introductionmentioning

confidence: 99%

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

Costa

Alves

Carneiro

et al. 2019

IJMS

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“…Although ELISA showed high specificity when comparing healthy individuals, the results in this review confirmed that this does not occur when the individuals examined have other diseases, especially Chagas’ disease and paracoccidioidomycosis. The same can occur with cases of VL and Leprosy [60] and with the adoption of IFR and cytometry. Thus, it is essential that specific antigens be developed and that, in the case of application of these tests in clinical practice, a differential diagnosis be made [7].…”

Section: Discussionmentioning

confidence: 99%

Accuracy of diagnostic tests for American tegumentary leishmaniasis: a systematic literature review with meta‐analyses

Pena

Belo

Xavier‐Júnior

et al. 2020

Tropical Med Int Health

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objectives To analyse the accuracy of American tegumentary leishmaniasis (ATL) diagnostic methods and evaluate the quality of the existing publications by means of a systematic review. methods Diagnostic tests evaluated in at least two studies with common reference standards were included in the sensitivity and/or specificity meta-analyses. Quality and susceptibility to bias were analysed using the QUADAS-2 and STARD tools. results The title and abstract of 3387 publications were evaluated after deduplication resulting from database searches. Thirty-eight studies were included in the review, and 26 of them had results inserted in meta-analyses. The diagnostic methods with the highest pooled sensitivity values were ELISA, polymerase chain reaction (PCR), indirect immunofluorescence reaction and Montenegro's intradermal reaction. Cytometry was assessed in only two studies and presented 100% sensitivity in both. Smear slide microscopy and histopathology showed low pooled values of sensitivity. For specificity, the highest pooled values were identified for PCR. High values were also identified for ELISA, except for studies in which the reference standard for defining negative participants included individuals with Chagas' disease or paracoccidioidomycosis, which also occurred for cytometry. IFR had lower specificities than ELISA. There was a predominance of case-control designs of phase 1 or 2 and only four studies were strongly recommended as evidence generators. Several reference standards were adopted, and different methods were assessed in a small number of studies. conclusion PCR showed the highest accuracy for the diagnosis of ATL, and its use should be encouraged in clinical practice. ELISA is recommended for the screening of suspected individuals, but the possibility of cross-reactions should be considered. New validation studies for the tests evaluated in few publications and studies of phase 3 with appropriate methods are needed.

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“…Both antigens showed good performance, however, rLb6H showed greater reactivity with L. ( L. ) amazonensis , followed by L. ( V. ) braziliensis and L. ( V. ) guyanensis . Other recombinant proteins, such as LiHypA, SMP-3, CcOx, HRF, rLiHyL, and others, were evaluated and presented good results, with some studies observing good sensitivity and specificity for both CL and VL [ 187 , 193 , 194 , 195 ]. In general, these recombinant proteins have higher sensitivity and specificity than the soluble Leishmania antigen extract, but they do not discriminate CL and VL infection.…”

Section: Immunological Methods For Diagnosis Of Leishmaniasismentioning

confidence: 99%

Laboratory Diagnosis of Cutaneous and Visceral Leishmaniasis: Current and Future Methods

et al. 2020

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Leishmaniasis is a neglected tropical disease with two main clinical forms: cutaneous and visceral leishmaniasis. Diagnosis of leishmaniasis is still a challenge, concerning the detection and correct identification of the species of the parasite, mainly in endemic areas where the absence of appropriate resources is still a problem. Most accessible methods for diagnosis, particularly in these areas, do not include the identification of each one of more than 20 species responsible for the disease. Here, we summarize the main methods used for the detection and identification of leishmaniasis that can be performed by demonstration of the parasite in biological samples from the patient through microscopic examination, by in vitro culture or animal inoculation; by molecular methods through the detection of parasite DNA; or by immunological methods through the detection of parasite antigens that may be present in urine or through the detection of specific antibodies against the parasite. Potential new methods that can be applied for laboratory diagnosis of leishmaniasis are also discussed.

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Potential application of small myristoylated protein-3 evaluated as recombinant antigen and a synthetic peptide containing its linear B-cell epitope for the serodiagnosis of canine visceral and human tegumentary leishmaniasis

Cited by 16 publications

References 58 publications

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

Accuracy of diagnostic tests for American tegumentary leishmaniasis: a systematic literature review with meta‐analyses

Laboratory Diagnosis of Cutaneous and Visceral Leishmaniasis: Current and Future Methods

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