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Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope
BACKGROUND: Leishmaniasis is one of the most important zoonotic diseases spread in Latin America. Since many species are involved in dog infection with different clinical manifestations, the development of specific diagnostic tests is mandatory for more accurate disease control and vaccine strategie...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466330/ https://www.ncbi.nlm.nih.gov/pubmed/28557986 http://dx.doi.org/10.1371/journal.pntd.0005562 |
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author | Mendes, Thais Melo Roma, Eric Henrique Costal-Oliveira, Fernanda Dhom-Lemos, Lucas de Carvalho Toledo-Machado, Cristina Monerat Bruna-Romero, Oscar Bartholomeu, Daniella Castanheiras Fujiwara, Ricardo Toshio Chávez-Olórtegui, Carlos |
author_facet | Mendes, Thais Melo Roma, Eric Henrique Costal-Oliveira, Fernanda Dhom-Lemos, Lucas de Carvalho Toledo-Machado, Cristina Monerat Bruna-Romero, Oscar Bartholomeu, Daniella Castanheiras Fujiwara, Ricardo Toshio Chávez-Olórtegui, Carlos |
author_sort | Mendes, Thais Melo |
collection | PubMed |
description | BACKGROUND: Leishmaniasis is one of the most important zoonotic diseases spread in Latin America. Since many species are involved in dog infection with different clinical manifestations, the development of specific diagnostic tests is mandatory for more accurate disease control and vaccine strategies. METHODOLOGY/PRINCIPAL FINDINGS: Seventy-five 15-mer peptides covering the sequence of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) protein were prepared by Spot synthesis. Membrane-bound peptides immunoreactivity with sera from dogs immunized with recLdVFA2 and with a specific anti-recLdVFA2 monoclonal antibody allowed mapping of continuous B-cell epitopes. Five epitopes corresponding to the N-terminal region of recLdVFA2 (MKIRSVRPLVVLLVC, RSVRPLVVLLVCVAA, RPLVVLLVCVAAVLA, VVLLVCVAAVLALSA and LVCVAAVLALSASAE, region 1–28) and one located within the repetitive units (PLSVGPQAVGLSVG, regions 67–81 and 122–135) were identified. A 34-mer recLdVFA2-derived bi-epitope containing the sequence MKIRSVRPLVVLLVC linked to PLSVGPQAVGLSVG by a Gly-Gly spacer was chemically synthesized in its soluble form. The synthetic bi-epitope was used as antigen to coat ELISA plates and assayed with dog sera for in vitro diagnosis of canine visceral leishmaniasis (CVL). The assay proved to be highly sensitive (98%) and specific (99%). CONCLUSIONS/SIGNIFICANCE: Our work suggests that synthetic peptide-based ELISA strategy may be useful for the development of a sensitive and highly specific serodiagnosis for CVL or other parasitic diseases. |
format | Online Article Text |
id | pubmed-5466330 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-54663302017-06-22 Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope Mendes, Thais Melo Roma, Eric Henrique Costal-Oliveira, Fernanda Dhom-Lemos, Lucas de Carvalho Toledo-Machado, Cristina Monerat Bruna-Romero, Oscar Bartholomeu, Daniella Castanheiras Fujiwara, Ricardo Toshio Chávez-Olórtegui, Carlos PLoS Negl Trop Dis Research Article BACKGROUND: Leishmaniasis is one of the most important zoonotic diseases spread in Latin America. Since many species are involved in dog infection with different clinical manifestations, the development of specific diagnostic tests is mandatory for more accurate disease control and vaccine strategies. METHODOLOGY/PRINCIPAL FINDINGS: Seventy-five 15-mer peptides covering the sequence of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) protein were prepared by Spot synthesis. Membrane-bound peptides immunoreactivity with sera from dogs immunized with recLdVFA2 and with a specific anti-recLdVFA2 monoclonal antibody allowed mapping of continuous B-cell epitopes. Five epitopes corresponding to the N-terminal region of recLdVFA2 (MKIRSVRPLVVLLVC, RSVRPLVVLLVCVAA, RPLVVLLVCVAAVLA, VVLLVCVAAVLALSA and LVCVAAVLALSASAE, region 1–28) and one located within the repetitive units (PLSVGPQAVGLSVG, regions 67–81 and 122–135) were identified. A 34-mer recLdVFA2-derived bi-epitope containing the sequence MKIRSVRPLVVLLVC linked to PLSVGPQAVGLSVG by a Gly-Gly spacer was chemically synthesized in its soluble form. The synthetic bi-epitope was used as antigen to coat ELISA plates and assayed with dog sera for in vitro diagnosis of canine visceral leishmaniasis (CVL). The assay proved to be highly sensitive (98%) and specific (99%). CONCLUSIONS/SIGNIFICANCE: Our work suggests that synthetic peptide-based ELISA strategy may be useful for the development of a sensitive and highly specific serodiagnosis for CVL or other parasitic diseases. Public Library of Science 2017-05-30 /pmc/articles/PMC5466330/ /pubmed/28557986 http://dx.doi.org/10.1371/journal.pntd.0005562 Text en © 2017 Mendes et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Mendes, Thais Melo Roma, Eric Henrique Costal-Oliveira, Fernanda Dhom-Lemos, Lucas de Carvalho Toledo-Machado, Cristina Monerat Bruna-Romero, Oscar Bartholomeu, Daniella Castanheiras Fujiwara, Ricardo Toshio Chávez-Olórtegui, Carlos Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope |
title | Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope |
title_full | Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope |
title_fullStr | Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope |
title_full_unstemmed | Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope |
title_short | Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope |
title_sort | epitope mapping of recombinant leishmania donovani virulence factor a2 (recldvfa2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466330/ https://www.ncbi.nlm.nih.gov/pubmed/28557986 http://dx.doi.org/10.1371/journal.pntd.0005562 |
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