Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs.
Visceral leishmaniasis is the most severe form of leishmaniasis. Worldwide, approximately 20% of zoonotic human visceral leishmaniasis is caused by Leishmania infantum, also known as Leishmania chagasi in Latin America. Current diagnostic methods are not accurate enough to identify Leishmania-infect...
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ftdoajarticles:oai:doaj.org/article:429315f53992415e8d9b9bfb52692322 2023-05-15T15:11:55+02:00 Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs. Angélica Rosa Faria Luciano de Castro Veloso Wendel Coura-Vital Alexandre Barbosa Reis Leonardo Miranda Damasceno Ricardo T Gazzinelli Hélida M Andrade 2015-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0003429 https://doaj.org/article/429315f53992415e8d9b9bfb52692322 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC4287523?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0003429 https://doaj.org/article/429315f53992415e8d9b9bfb52692322 PLoS Neglected Tropical Diseases, Vol 9, Iss 1, p e3429 (2015) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2015 ftdoajarticles https://doi.org/10.1371/journal.pntd.0003429 2022-12-31T10:46:05Z Visceral leishmaniasis is the most severe form of leishmaniasis. Worldwide, approximately 20% of zoonotic human visceral leishmaniasis is caused by Leishmania infantum, also known as Leishmania chagasi in Latin America. Current diagnostic methods are not accurate enough to identify Leishmania-infected animals and may compromise the effectiveness of disease control. Therefore, we aimed to produce and test two recombinant multiepitope proteins as a means to improve and increase accuracy in the diagnosis of canine visceral leishmaniasis (CVL).Ten antigenic peptides were identified by CVL ELISA in previous work. In the current proposal, the coding sequences of these ten peptides were assembled into a synthetic gene. Furthermore, other twenty peptides were selected from work by our group where good B and T cell epitopes were mapped. The coding sequences of these peptides were also assembled into a synthetic gene. Both genes have been cloned and expressed in Escherichia coli, producing two multiepitope recombinant proteins, PQ10 and PQ20. These antigens have been used in CVL ELISA and were able to identify asymptomatic dogs (80%) more effectively than EIE-LVC kit, produced by Bio-Manguinhos (0%) and DPP kit (10%). Moreover, our recombinant proteins presented an early detection (before PCR) of infected dogs, with positivities ranging from 23% to 65%, depending on the phase of infection in which sera were acquired.Our study shows that ELISA using the multiepitope proteins PQ10 and PQ20 has great potential in early CVL diagnosis. The use of these proteins in other methodologies, such as immunochromatographic tests, could be beneficial mainly for the detection of asymptomatic dogs. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Eie ENVELOPE(7.983,7.983,63.033,63.033) PLoS Neglected Tropical Diseases 9 1 e3429 |
institution |
Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
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 Angélica Rosa Faria Luciano de Castro Veloso Wendel Coura-Vital Alexandre Barbosa Reis Leonardo Miranda Damasceno Ricardo T Gazzinelli Hélida M Andrade Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Visceral leishmaniasis is the most severe form of leishmaniasis. Worldwide, approximately 20% of zoonotic human visceral leishmaniasis is caused by Leishmania infantum, also known as Leishmania chagasi in Latin America. Current diagnostic methods are not accurate enough to identify Leishmania-infected animals and may compromise the effectiveness of disease control. Therefore, we aimed to produce and test two recombinant multiepitope proteins as a means to improve and increase accuracy in the diagnosis of canine visceral leishmaniasis (CVL).Ten antigenic peptides were identified by CVL ELISA in previous work. In the current proposal, the coding sequences of these ten peptides were assembled into a synthetic gene. Furthermore, other twenty peptides were selected from work by our group where good B and T cell epitopes were mapped. The coding sequences of these peptides were also assembled into a synthetic gene. Both genes have been cloned and expressed in Escherichia coli, producing two multiepitope recombinant proteins, PQ10 and PQ20. These antigens have been used in CVL ELISA and were able to identify asymptomatic dogs (80%) more effectively than EIE-LVC kit, produced by Bio-Manguinhos (0%) and DPP kit (10%). Moreover, our recombinant proteins presented an early detection (before PCR) of infected dogs, with positivities ranging from 23% to 65%, depending on the phase of infection in which sera were acquired.Our study shows that ELISA using the multiepitope proteins PQ10 and PQ20 has great potential in early CVL diagnosis. The use of these proteins in other methodologies, such as immunochromatographic tests, could be beneficial mainly for the detection of asymptomatic dogs. |
format |
Article in Journal/Newspaper |
author |
Angélica Rosa Faria Luciano de Castro Veloso Wendel Coura-Vital Alexandre Barbosa Reis Leonardo Miranda Damasceno Ricardo T Gazzinelli Hélida M Andrade |
author_facet |
Angélica Rosa Faria Luciano de Castro Veloso Wendel Coura-Vital Alexandre Barbosa Reis Leonardo Miranda Damasceno Ricardo T Gazzinelli Hélida M Andrade |
author_sort |
Angélica Rosa Faria |
title |
Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs. |
title_short |
Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs. |
title_full |
Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs. |
title_fullStr |
Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs. |
title_full_unstemmed |
Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs. |
title_sort |
novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2015 |
url |
https://doi.org/10.1371/journal.pntd.0003429 https://doaj.org/article/429315f53992415e8d9b9bfb52692322 |
long_lat |
ENVELOPE(7.983,7.983,63.033,63.033) |
geographic |
Arctic Eie |
geographic_facet |
Arctic Eie |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 9, Iss 1, p e3429 (2015) |
op_relation |
http://europepmc.org/articles/PMC4287523?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0003429 https://doaj.org/article/429315f53992415e8d9b9bfb52692322 |
op_doi |
https://doi.org/10.1371/journal.pntd.0003429 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
9 |
container_issue |
1 |
container_start_page |
e3429 |
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1766342697351643136 |