PMID- 21111760
OWN - NLM
STAT- MEDLINE
DCOM- 20110404
LR  - 20191210
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 171
IP  - 1
DP  - 2011 Jan
TI  - Development of a highly sensitive real-time RT-PCR protocol for the detection of 
      Classical swine fever virus independent of the 5' untranslated region.
PG  - 314-7
LID - 10.1016/j.jviromet.2010.11.014 [doi]
AB  - Classical swine fever (CSF) is one of the most severe diseases of pigs, and can 
      cause immense economic losses. Real-time reverse transcription polymerase chain 
      reaction (rRT-PCR) can be used as a sensitive and specific method for detection 
      of Classical swine fever virus (CSFV). Different published protocols are used 
      routinely for CSFV diagnosis. However, almost all these systems use the highly 
      conserved 5' untranslated region (5'UTR) of the CSFV genome as template. For a 
      reliable diagnosis in outbreaks, a confirmatory assay that amplifies a different 
      genome region is advisable. In this study a new CSFV specific rRT-PCR system 
      using the NS5A region as template is described. The assay is multiplexed with a 
      beta-actin detection system that is used as an internal control in a single tube 
      assay. The system was validated using recent European CSFV field isolates, 
      dilution series of an in vitro transcribed RNA standard, and a panel of RNAs 
      representing all available Pestivirus species and genotypes. It was shown that 
      the new assay allows reliable detection of CSFV genomes independent of the 5'UTR 
      region. It presents a very useful diagnostic tool, also allowing a 'double check' 
      approach to rule out 5'UTR amplicon contaminations.
CI  - Copyright (c) 2010 Elsevier B.V. All rights reserved.
FAU - Leifer, Immanuel
AU  - Leifer I
AD  - Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Greifswald-Insel 
      Riems, Germany.
FAU - Blome, Sandra
AU  - Blome S
FAU - Beer, Martin
AU  - Beer M
FAU - Hoffmann, Bernd
AU  - Hoffmann B
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Validation Study
DEP - 20101125
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (5' Untranslated Regions)
RN  - 0 (DNA Primers)
RN  - 0 (NS5 protein, flavivirus)
RN  - 0 (Viral Nonstructural Proteins)
SB  - IM
MH  - 5' Untranslated Regions
MH  - Animals
MH  - Classical Swine Fever/*diagnosis/virology
MH  - Classical Swine Fever Virus/genetics/*isolation & purification
MH  - DNA Primers/genetics
MH  - Reference Standards
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods/standards
MH  - Sensitivity and Specificity
MH  - Swine
MH  - Viral Nonstructural Proteins/*genetics
MH  - Virology/*methods/standards
EDAT- 2010/11/30 06:00
MHDA- 2011/04/05 06:00
CRDT- 2010/11/30 06:00
PHST- 2010/07/10 00:00 [received]
PHST- 2010/11/15 00:00 [revised]
PHST- 2010/11/18 00:00 [accepted]
PHST- 2010/11/30 06:00 [entrez]
PHST- 2010/11/30 06:00 [pubmed]
PHST- 2011/04/05 06:00 [medline]
AID - S0166-0934(10)00413-1 [pii]
AID - 10.1016/j.jviromet.2010.11.014 [doi]
PST - ppublish
SO  - J Virol Methods. 2011 Jan;171(1):314-7. doi: 10.1016/j.jviromet.2010.11.014. Epub 
      2010 Nov 25.