Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks

Detalhes bibliográficos
Autor(a) principal: Radulović,Ž.
Data de Publicação: 2010
Outros Autores: Milutinović,M., Tomanović,S., Mulenga,A.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Arquivo brasileiro de medicina veterinária e zootecnia (Online)
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352010000400015
Resumo: A reverse transcriptase - polymerase chain reaction based assay for Borrelia species detection in ticks was developed. The method was based on amplification of 552 nucleotide bases long sequence of 16S rRNA, targeted by Borrelia specific primers. In the present study, total RNA extracted from Ixodes ricinus ticks was used as template. The results showed higher sensitivity for Borrelia detection as compared to standard dark-field microscopy. Method specificity was confirmed by cloning and sequencing of obtained 552 base pairs long amplicons. Phylogenetic analysis of obtained sequences showed that they belong to B. lusitaniae and B. afzelii genospecies. RT-PCR based method presented in this paper could be very useful as a screening test for detecting pathogen presence, especially when in investigations is required extraction of total RNA from ticks.
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spelling Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticksBorreliaRT-PCR16S rRNAIxodes ricinusA reverse transcriptase - polymerase chain reaction based assay for Borrelia species detection in ticks was developed. The method was based on amplification of 552 nucleotide bases long sequence of 16S rRNA, targeted by Borrelia specific primers. In the present study, total RNA extracted from Ixodes ricinus ticks was used as template. The results showed higher sensitivity for Borrelia detection as compared to standard dark-field microscopy. Method specificity was confirmed by cloning and sequencing of obtained 552 base pairs long amplicons. Phylogenetic analysis of obtained sequences showed that they belong to B. lusitaniae and B. afzelii genospecies. RT-PCR based method presented in this paper could be very useful as a screening test for detecting pathogen presence, especially when in investigations is required extraction of total RNA from ticks.Universidade Federal de Minas Gerais, Escola de Veterinária2010-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352010000400015Arquivo Brasileiro de Medicina Veterinária e Zootecnia v.62 n.4 2010reponame:Arquivo brasileiro de medicina veterinária e zootecnia (Online)instname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMG10.1590/S0102-09352010000400015info:eu-repo/semantics/openAccessRadulović,Ž.Milutinović,M.Tomanović,S.Mulenga,A.eng2011-02-02T00:00:00Zoai:scielo:S0102-09352010000400015Revistahttps://www.scielo.br/j/abmvz/PUBhttps://old.scielo.br/oai/scielo-oai.phpjournal@vet.ufmg.br||abmvz.artigo@abmvz.org.br1678-41620102-0935opendoar:2011-02-02T00:00Arquivo brasileiro de medicina veterinária e zootecnia (Online) - Universidade Federal de Minas Gerais (UFMG)false
dc.title.none.fl_str_mv Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks
title Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks
spellingShingle Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks
Radulović,Ž.
Borrelia
RT-PCR
16S rRNA
Ixodes ricinus
title_short Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks
title_full Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks
title_fullStr Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks
title_full_unstemmed Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks
title_sort Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks
author Radulović,Ž.
author_facet Radulović,Ž.
Milutinović,M.
Tomanović,S.
Mulenga,A.
author_role author
author2 Milutinović,M.
Tomanović,S.
Mulenga,A.
author2_role author
author
author
dc.contributor.author.fl_str_mv Radulović,Ž.
Milutinović,M.
Tomanović,S.
Mulenga,A.
dc.subject.por.fl_str_mv Borrelia
RT-PCR
16S rRNA
Ixodes ricinus
topic Borrelia
RT-PCR
16S rRNA
Ixodes ricinus
description A reverse transcriptase - polymerase chain reaction based assay for Borrelia species detection in ticks was developed. The method was based on amplification of 552 nucleotide bases long sequence of 16S rRNA, targeted by Borrelia specific primers. In the present study, total RNA extracted from Ixodes ricinus ticks was used as template. The results showed higher sensitivity for Borrelia detection as compared to standard dark-field microscopy. Method specificity was confirmed by cloning and sequencing of obtained 552 base pairs long amplicons. Phylogenetic analysis of obtained sequences showed that they belong to B. lusitaniae and B. afzelii genospecies. RT-PCR based method presented in this paper could be very useful as a screening test for detecting pathogen presence, especially when in investigations is required extraction of total RNA from ticks.
publishDate 2010
dc.date.none.fl_str_mv 2010-08-01
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=S0102-09352010000400015
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352010000400015
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S0102-09352010000400015
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 Universidade Federal de Minas Gerais, Escola de Veterinária
publisher.none.fl_str_mv Universidade Federal de Minas Gerais, Escola de Veterinária
dc.source.none.fl_str_mv Arquivo Brasileiro de Medicina Veterinária e Zootecnia v.62 n.4 2010
reponame:Arquivo brasileiro de medicina veterinária e zootecnia (Online)
instname:Universidade Federal de Minas Gerais (UFMG)
instacron:UFMG
instname_str Universidade Federal de Minas Gerais (UFMG)
instacron_str UFMG
institution UFMG
reponame_str Arquivo brasileiro de medicina veterinária e zootecnia (Online)
collection Arquivo brasileiro de medicina veterinária e zootecnia (Online)
repository.name.fl_str_mv Arquivo brasileiro de medicina veterinária e zootecnia (Online) - Universidade Federal de Minas Gerais (UFMG)
repository.mail.fl_str_mv journal@vet.ufmg.br||abmvz.artigo@abmvz.org.br
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