Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A

Detalhes bibliográficos
Autor(a) principal: Ferreira,Helena Lage
Data de Publicação: 2009
Outros Autores: Spilki,Fernando Rosado, Santos,Márcia Mercês Aparecida Bianchi dos, Almeida,Renata Servan de, Arns,Clarice Weis
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Ciência Rural
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782009000500022
Resumo: Avian metapneumovirus (AMPV) belongs to Metapneumovirus genus of Paramyxoviridae family. Virus isolation, serology, and detection of genomic RNA are used as diagnostic methods for AMPV. The aim of the present study was to compare the detection of six subgroup A AMPV isolates (AMPV/A) viral RNA by using different conventional and real time RT-PCR methods. Two new RT-PCR tests and two real time RT-PCR tests, both detecting fusion (F) gene and nucleocapsid (N) gene were compared with an established test for the attachment (G) gene. All the RT-PCR tested assays were able to detect the AMPV/A. The lower detection limits were observed using the N-, F- based RRT-PCR and F-based conventional RT-PCR (10(0.3) to 10¹ TCID50 mL-1). The present study suggests that the conventional F-based RT-PCR presented similar detection limit when compared to N- and F-based RRT-PCR and they can be successfully used for AMPV/A detection.
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spelling Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype Aavian metapneumovirusG, F, N genesreal time RT-PCRRT-PCRAvian metapneumovirus (AMPV) belongs to Metapneumovirus genus of Paramyxoviridae family. Virus isolation, serology, and detection of genomic RNA are used as diagnostic methods for AMPV. The aim of the present study was to compare the detection of six subgroup A AMPV isolates (AMPV/A) viral RNA by using different conventional and real time RT-PCR methods. Two new RT-PCR tests and two real time RT-PCR tests, both detecting fusion (F) gene and nucleocapsid (N) gene were compared with an established test for the attachment (G) gene. All the RT-PCR tested assays were able to detect the AMPV/A. The lower detection limits were observed using the N-, F- based RRT-PCR and F-based conventional RT-PCR (10(0.3) to 10¹ TCID50 mL-1). The present study suggests that the conventional F-based RT-PCR presented similar detection limit when compared to N- and F-based RRT-PCR and they can be successfully used for AMPV/A detection.Universidade Federal de Santa Maria2009-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782009000500022Ciência Rural v.39 n.5 2009reponame:Ciência Ruralinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM10.1590/S0103-84782009005000057info:eu-repo/semantics/openAccessFerreira,Helena LageSpilki,Fernando RosadoSantos,Márcia Mercês Aparecida Bianchi dosAlmeida,Renata Servan deArns,Clarice Weiseng2009-07-28T00:00:00ZRevista
dc.title.none.fl_str_mv Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A
title Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A
spellingShingle Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A
Ferreira,Helena Lage
avian metapneumovirus
G, F, N genes
real time RT-PCR
RT-PCR
title_short Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A
title_full Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A
title_fullStr Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A
title_full_unstemmed Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A
title_sort Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A
author Ferreira,Helena Lage
author_facet Ferreira,Helena Lage
Spilki,Fernando Rosado
Santos,Márcia Mercês Aparecida Bianchi dos
Almeida,Renata Servan de
Arns,Clarice Weis
author_role author
author2 Spilki,Fernando Rosado
Santos,Márcia Mercês Aparecida Bianchi dos
Almeida,Renata Servan de
Arns,Clarice Weis
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Ferreira,Helena Lage
Spilki,Fernando Rosado
Santos,Márcia Mercês Aparecida Bianchi dos
Almeida,Renata Servan de
Arns,Clarice Weis
dc.subject.por.fl_str_mv avian metapneumovirus
G, F, N genes
real time RT-PCR
RT-PCR
topic avian metapneumovirus
G, F, N genes
real time RT-PCR
RT-PCR
description Avian metapneumovirus (AMPV) belongs to Metapneumovirus genus of Paramyxoviridae family. Virus isolation, serology, and detection of genomic RNA are used as diagnostic methods for AMPV. The aim of the present study was to compare the detection of six subgroup A AMPV isolates (AMPV/A) viral RNA by using different conventional and real time RT-PCR methods. Two new RT-PCR tests and two real time RT-PCR tests, both detecting fusion (F) gene and nucleocapsid (N) gene were compared with an established test for the attachment (G) gene. All the RT-PCR tested assays were able to detect the AMPV/A. The lower detection limits were observed using the N-, F- based RRT-PCR and F-based conventional RT-PCR (10(0.3) to 10¹ TCID50 mL-1). The present study suggests that the conventional F-based RT-PCR presented similar detection limit when compared to N- and F-based RRT-PCR and they can be successfully used for AMPV/A detection.
publishDate 2009
dc.date.none.fl_str_mv 2009-08-01
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=S0103-84782009000500022
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782009000500022
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S0103-84782009005000057
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 Santa Maria
publisher.none.fl_str_mv Universidade Federal de Santa Maria
dc.source.none.fl_str_mv Ciência Rural v.39 n.5 2009
reponame:Ciência Rural
instname:Universidade Federal de Santa Maria (UFSM)
instacron:UFSM
instname_str Universidade Federal de Santa Maria (UFSM)
instacron_str UFSM
institution UFSM
reponame_str Ciência Rural
collection Ciência Rural
repository.name.fl_str_mv
repository.mail.fl_str_mv
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