Bluetongue virus detection by two real-time RT-qPCRs targeting two different genomic segments.
J Virol Methods
; 140(1-2): 115-23, 2007 Mar.
Article
em En
| MEDLINE
| ID: mdl-17196266
ABSTRACT
The detection of the bluetongue virus (BTV) by conventional methods is especially difficult and labour-intensive. Molecular diagnosis is also complex because of the high genetic diversity between and within the 24 serotypes of BTV. In the present study, two laboratories joined forces to develop and validate two new RT-qPCRs detecting and amplifying BTV segments 1 and 5. The 2 assays detect strains from all 24 serotypes. They both have a detection limit of 0.01 ECE50 and all 114 samples from BTV-free goats, sheep and cattle were negative. The two assays resulted in similar C(t) values when testing biological samples collected in sheep infected experimentally with a field strain of BTV from the Mediterranean basin. On average, the C(t) values obtained with the 2 methods applied to the 24 serotypes were not significantly different from each other, but some moderate to high differences were seen with a few strains. Therefore these two methods are complementary and could be used in parallel to confirm the diagnosis of a possible new introduction of BTV. An RT-qPCR amplifying a fragment of the beta-actin mRNA was also developed and validated as internal control for the bluetongue specific assays. The three assays described allow a reliable and rapid detection of BTV.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Doenças dos Ovinos
/
Genoma Viral
/
Bluetongue
/
Vírus Bluetongue
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
J Virol Methods
Ano de publicação:
2007
Tipo de documento:
Article
País de afiliação:
Bélgica