Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus

Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus

Andrés, Damián F. de [et. al]. 7 p., 3 tables, 3 figures and bibliography

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Main Authors: Andrés, Damián F. de, Amorena Zabalza, Beatriz
Format: artículo biblioteca
Language:English
Published: Elsevier 2010-05
Subjects:Lentivirus, Visna/maedi virus, EV1, Real-time PCR, Proviral load, Viral transcript,
Online Access:http://hdl.handle.net/10261/48416
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spelling dig-idab-es-10261-484162016-09-07T11:46:37Z Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus Andrés, Damián F. de Amorena Zabalza, Beatriz Lentivirus Visna/maedi virus EV1 Real-time PCR Proviral load Viral transcript Andrés, Damián F. de [et. al]. 7 p., 3 tables, 3 figures and bibliography The aim of this study was the development of gag and pol dual labelled probe real-time PCR and RT PCR assays to quantify the proviral load and the transcripts of the British Visna/maedi virus EV1 strain. Primers and probes were chosen based on the consensus sequences of gag and pol clones representative of EV1 genetic variants. Both PCRs had a detection limit of 3 copies of target gene, with a linearity over 6 orders of magnitude. The performances of the two PCRs in vivo were evaluated and compared on a panel of DNAs extracted from blood of sheep infected experimentally with EV1. The pol assay detected in most cases lower numbers of viral molecules than gag assay, yielding some false negative results. The gag real-time RT PCR had a detection limit of 100 RNA molecules with a linearity over 5 orders of magnitude. This did not result in a lower performance of the RT PCR compared to the PCR in cells permissive for virus replication, which contain higher numbers of viral transcripts than proviral genomes. The real-time assays developed in this study, particularly the gag assay, provide a sensitive tool which can be used to quantify the viral load in experimental infections. This study was supported by grants from European Union (QLK2-CT-2002-00617). Peer reviewed 2012-04-17T12:20:43Z 2012-04-17T12:20:43Z 2010-05 artículo http://purl.org/coar/resource_type/c_6501 Journal of Virological Methods 165(2): 161-167 (2010) 0166-0934 http://hdl.handle.net/10261/48416 10.1016/j.jviromet.2010.01.013 1879-0984 en http://dx.doi.org/10.1016/j.jviromet.2010.01.013 none Elsevier
institution IDAB ES
collection DSpace
country España
countrycode ES
component Bibliográfico
access En linea
databasecode dig-idab-es
tag biblioteca
region Europa del Sur
libraryname Biblioteca del IDAB España
language English
topic Lentivirus
Visna/maedi virus
EV1
Real-time PCR
Proviral load
Viral transcript
Lentivirus
Visna/maedi virus
EV1
Real-time PCR
Proviral load
Viral transcript
spellingShingle Lentivirus
Visna/maedi virus
EV1
Real-time PCR
Proviral load
Viral transcript
Lentivirus
Visna/maedi virus
EV1
Real-time PCR
Proviral load
Viral transcript
Andrés, Damián F. de
Amorena Zabalza, Beatriz
Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus
description Andrés, Damián F. de [et. al]. 7 p., 3 tables, 3 figures and bibliography
format artículo
topic_facet Lentivirus
Visna/maedi virus
EV1
Real-time PCR
Proviral load
Viral transcript
author Andrés, Damián F. de
Amorena Zabalza, Beatriz
author_facet Andrés, Damián F. de
Amorena Zabalza, Beatriz
author_sort Andrés, Damián F. de
title Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus
title_short Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus
title_full Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus
title_fullStr Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus
title_full_unstemmed Development and comparison of strain specific gag and pol real-time PCR assays for the detection of Visna/maedi virus
title_sort development and comparison of strain specific gag and pol real-time pcr assays for the detection of visna/maedi virus
publisher Elsevier
publishDate 2010-05
url http://hdl.handle.net/10261/48416
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