Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Revista Brasileira de Parasitologia Veterinária (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612020000300304 |
Resumo: | Abstract Ehrlichia canis is the main etiological agent of canine monocytic ehrlichiosis (CME), a globally canine infectious disease. In Brazil, CME is considered to be endemic, and its prevalence can reach 65% in some states. The diagnosis of ehrlichiosis is important for treatment and epidemiological purposes. The E. canis TRP36 (Tandem Repeat Protein) protein elicits the earliest acute-phase antibody response observed during the course of the disease. This study aimed to generate the recombinant TRP36 protein from E. canis São Paulo strain and to evaluate its potential as a tool for the serologic diagnosis of CME. The E. canis São Paulo isolate was cultivated in DH82 lineage cells, and its genomic DNA was obtained. The bacterial DNA fragment encoding the entire ORF of TRP36 was cloned into the pBAD/Thio-TOPO vector and transformed into Escherichia coli DH10B competent cells with the trp36-bearing plasmid for protein expression. To evaluate the protein antigenicity, 16 canine serum samples were previously tested (by PCR and the commercial SNAP®4Dx® serological test). The results were in accordance with the SNAP®4Dx® test. Experiments using this recombinant protein as an antigen, targeting the development of a serologic test based on ELISA methodology, are the next step to produce a reliable, affordable and useful diagnostic tool for CME in Brazil. |
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Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic testsEhrlichia canisTRP36 proteinepidemiologydiagnosticserologic testsAbstract Ehrlichia canis is the main etiological agent of canine monocytic ehrlichiosis (CME), a globally canine infectious disease. In Brazil, CME is considered to be endemic, and its prevalence can reach 65% in some states. The diagnosis of ehrlichiosis is important for treatment and epidemiological purposes. The E. canis TRP36 (Tandem Repeat Protein) protein elicits the earliest acute-phase antibody response observed during the course of the disease. This study aimed to generate the recombinant TRP36 protein from E. canis São Paulo strain and to evaluate its potential as a tool for the serologic diagnosis of CME. The E. canis São Paulo isolate was cultivated in DH82 lineage cells, and its genomic DNA was obtained. The bacterial DNA fragment encoding the entire ORF of TRP36 was cloned into the pBAD/Thio-TOPO vector and transformed into Escherichia coli DH10B competent cells with the trp36-bearing plasmid for protein expression. To evaluate the protein antigenicity, 16 canine serum samples were previously tested (by PCR and the commercial SNAP®4Dx® serological test). The results were in accordance with the SNAP®4Dx® test. Experiments using this recombinant protein as an antigen, targeting the development of a serologic test based on ELISA methodology, are the next step to produce a reliable, affordable and useful diagnostic tool for CME in Brazil.Colégio Brasileiro de Parasitologia Veterinária2020-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612020000300304Revista Brasileira de Parasitologia Veterinária v.29 n.3 2020reponame:Revista Brasileira de Parasitologia Veterinária (Online)instname:Colégio Brasileiro de Parasitologia Veterinária (CBPV)instacron:CBPV10.1590/s1984-29612020051info:eu-repo/semantics/openAccessMedeiros,Miguel Ângelo da SilvaSilva,Maria Helena daMatta,Maria Adelaide do ValleFerreira,Eliane de OliveiraMachado,Sérgio LisboaSoares,João FábioLabruna,Marcelo BahiaToma,Helena KeikoXavier,Márcia de SouzaMeirelles,Maria de Nazareth Silveira Leal deAlmosny,Nádia Regina Pereiraeng2020-07-31T00:00:00Zoai:scielo:S1984-29612020000300304Revistahttp://www.scielo.br/scielo.php?script=sci_serial&lng=pt&pid=1984-2961https://old.scielo.br/oai/scielo-oai.php||zacariascbpv@fcav.unesp.br1984-29610103-846Xopendoar:2020-07-31T00:00Revista Brasileira de Parasitologia Veterinária (Online) - Colégio Brasileiro de Parasitologia Veterinária (CBPV)false |
dc.title.none.fl_str_mv |
Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests |
title |
Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests |
spellingShingle |
Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests Medeiros,Miguel Ângelo da Silva Ehrlichia canis TRP36 protein epidemiology diagnostic serologic tests |
title_short |
Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests |
title_full |
Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests |
title_fullStr |
Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests |
title_full_unstemmed |
Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests |
title_sort |
Expression and antigenic analysis of the recombinant TRP36 protein from Ehrlichia canis São Paulo strain for serologic tests |
author |
Medeiros,Miguel Ângelo da Silva |
author_facet |
Medeiros,Miguel Ângelo da Silva Silva,Maria Helena da Matta,Maria Adelaide do Valle Ferreira,Eliane de Oliveira Machado,Sérgio Lisboa Soares,João Fábio Labruna,Marcelo Bahia Toma,Helena Keiko Xavier,Márcia de Souza Meirelles,Maria de Nazareth Silveira Leal de Almosny,Nádia Regina Pereira |
author_role |
author |
author2 |
Silva,Maria Helena da Matta,Maria Adelaide do Valle Ferreira,Eliane de Oliveira Machado,Sérgio Lisboa Soares,João Fábio Labruna,Marcelo Bahia Toma,Helena Keiko Xavier,Márcia de Souza Meirelles,Maria de Nazareth Silveira Leal de Almosny,Nádia Regina Pereira |
author2_role |
author author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Medeiros,Miguel Ângelo da Silva Silva,Maria Helena da Matta,Maria Adelaide do Valle Ferreira,Eliane de Oliveira Machado,Sérgio Lisboa Soares,João Fábio Labruna,Marcelo Bahia Toma,Helena Keiko Xavier,Márcia de Souza Meirelles,Maria de Nazareth Silveira Leal de Almosny,Nádia Regina Pereira |
dc.subject.por.fl_str_mv |
Ehrlichia canis TRP36 protein epidemiology diagnostic serologic tests |
topic |
Ehrlichia canis TRP36 protein epidemiology diagnostic serologic tests |
description |
Abstract Ehrlichia canis is the main etiological agent of canine monocytic ehrlichiosis (CME), a globally canine infectious disease. In Brazil, CME is considered to be endemic, and its prevalence can reach 65% in some states. The diagnosis of ehrlichiosis is important for treatment and epidemiological purposes. The E. canis TRP36 (Tandem Repeat Protein) protein elicits the earliest acute-phase antibody response observed during the course of the disease. This study aimed to generate the recombinant TRP36 protein from E. canis São Paulo strain and to evaluate its potential as a tool for the serologic diagnosis of CME. The E. canis São Paulo isolate was cultivated in DH82 lineage cells, and its genomic DNA was obtained. The bacterial DNA fragment encoding the entire ORF of TRP36 was cloned into the pBAD/Thio-TOPO vector and transformed into Escherichia coli DH10B competent cells with the trp36-bearing plasmid for protein expression. To evaluate the protein antigenicity, 16 canine serum samples were previously tested (by PCR and the commercial SNAP®4Dx® serological test). The results were in accordance with the SNAP®4Dx® test. Experiments using this recombinant protein as an antigen, targeting the development of a serologic test based on ELISA methodology, are the next step to produce a reliable, affordable and useful diagnostic tool for CME in Brazil. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-01-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612020000300304 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612020000300304 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/s1984-29612020051 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Colégio Brasileiro de Parasitologia Veterinária |
publisher.none.fl_str_mv |
Colégio Brasileiro de Parasitologia Veterinária |
dc.source.none.fl_str_mv |
Revista Brasileira de Parasitologia Veterinária v.29 n.3 2020 reponame:Revista Brasileira de Parasitologia Veterinária (Online) instname:Colégio Brasileiro de Parasitologia Veterinária (CBPV) instacron:CBPV |
instname_str |
Colégio Brasileiro de Parasitologia Veterinária (CBPV) |
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CBPV |
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CBPV |
reponame_str |
Revista Brasileira de Parasitologia Veterinária (Online) |
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Revista Brasileira de Parasitologia Veterinária (Online) |
repository.name.fl_str_mv |
Revista Brasileira de Parasitologia Veterinária (Online) - Colégio Brasileiro de Parasitologia Veterinária (CBPV) |
repository.mail.fl_str_mv |
||zacariascbpv@fcav.unesp.br |
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1754208918766092288 |