Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope.
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.Seventy-fi...
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Online Access: | https://doi.org/10.1371/journal.pntd.0005562 https://doaj.org/article/3ae026b6a5c94ef1a1c7ea6a7d567f84 |
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ftdoajarticles:oai:doaj.org/article:3ae026b6a5c94ef1a1c7ea6a7d567f84 2023-05-15T15:08:52+02:00 Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope. Thais Melo Mendes Eric Henrique Roma Fernanda Costal-Oliveira Lucas de Carvalho Dhom-Lemos Cristina Monerat Toledo-Machado Oscar Bruna-Romero Daniella Castanheiras Bartholomeu Ricardo Toshio Fujiwara Carlos Chávez-Olórtegui 2017-05-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0005562 https://doaj.org/article/3ae026b6a5c94ef1a1c7ea6a7d567f84 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC5466330?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0005562 https://doaj.org/article/3ae026b6a5c94ef1a1c7ea6a7d567f84 PLoS Neglected Tropical Diseases, Vol 11, Iss 5, p e0005562 (2017) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2017 ftdoajarticles https://doi.org/10.1371/journal.pntd.0005562 2022-12-31T03:51:18Z 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.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%).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. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 11 5 e0005562 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Thais Melo Mendes Eric Henrique Roma Fernanda Costal-Oliveira Lucas de Carvalho Dhom-Lemos Cristina Monerat Toledo-Machado Oscar Bruna-Romero Daniella Castanheiras Bartholomeu Ricardo Toshio Fujiwara Carlos Chávez-Olórtegui Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
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.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%).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 |
Article in Journal/Newspaper |
author |
Thais Melo Mendes Eric Henrique Roma Fernanda Costal-Oliveira Lucas de Carvalho Dhom-Lemos Cristina Monerat Toledo-Machado Oscar Bruna-Romero Daniella Castanheiras Bartholomeu Ricardo Toshio Fujiwara Carlos Chávez-Olórtegui |
author_facet |
Thais Melo Mendes Eric Henrique Roma Fernanda Costal-Oliveira Lucas de Carvalho Dhom-Lemos Cristina Monerat Toledo-Machado Oscar Bruna-Romero Daniella Castanheiras Bartholomeu Ricardo Toshio Fujiwara Carlos Chávez-Olórtegui |
author_sort |
Thais Melo Mendes |
title |
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_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_sort |
epitope mapping of recombinant leishmania donovani virulence factor a2 (recldvfa2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2017 |
url |
https://doi.org/10.1371/journal.pntd.0005562 https://doaj.org/article/3ae026b6a5c94ef1a1c7ea6a7d567f84 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 11, Iss 5, p e0005562 (2017) |
op_relation |
http://europepmc.org/articles/PMC5466330?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0005562 https://doaj.org/article/3ae026b6a5c94ef1a1c7ea6a7d567f84 |
op_doi |
https://doi.org/10.1371/journal.pntd.0005562 |
container_title |
PLOS Neglected Tropical Diseases |
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11 |
container_issue |
5 |
container_start_page |
e0005562 |
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1766340145268654080 |