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...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
|
| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | http://europepmc.org/articles/PMC4287523?pdf=render |
| _version_ | 1818948188544958464 |
|---|---|
| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-20T08:42:50Z |
| format | Article |
| id | doaj.art-429315f53992415e8d9b9bfb52692322 |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-12-20T08:42:50Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-429315f53992415e8d9b9bfb526923222022-12-21T19:46:20ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352015-01-0191e342910.1371/journal.pntd.0003429Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs.Angélica Rosa FariaLuciano de Castro VelosoWendel Coura-VitalAlexandre Barbosa ReisLeonardo Miranda DamascenoRicardo T GazzinelliHélida M AndradeVisceral 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.http://europepmc.org/articles/PMC4287523?pdf=render |
| spellingShingle | 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. PLoS Neglected Tropical Diseases |
| title | 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_short | 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 |
| url | http://europepmc.org/articles/PMC4287523?pdf=render |
| work_keys_str_mv | AT angelicarosafaria novelrecombinantmultiepitopeproteinsforthediagnosisofasymptomaticleishmaniainfantuminfecteddogs AT lucianodecastroveloso novelrecombinantmultiepitopeproteinsforthediagnosisofasymptomaticleishmaniainfantuminfecteddogs AT wendelcouravital novelrecombinantmultiepitopeproteinsforthediagnosisofasymptomaticleishmaniainfantuminfecteddogs AT alexandrebarbosareis novelrecombinantmultiepitopeproteinsforthediagnosisofasymptomaticleishmaniainfantuminfecteddogs AT leonardomirandadamasceno novelrecombinantmultiepitopeproteinsforthediagnosisofasymptomaticleishmaniainfantuminfecteddogs AT ricardotgazzinelli novelrecombinantmultiepitopeproteinsforthediagnosisofasymptomaticleishmaniainfantuminfecteddogs AT helidamandrade novelrecombinantmultiepitopeproteinsforthediagnosisofasymptomaticleishmaniainfantuminfecteddogs |