Journal article
Quantitative multiplex assay for simultaneous detection and identification of Indiana and New Jersey serotypes of vesicular stomatitis virus
In order to establish a rapid and reliable system for the detection of vesicular stomatitis virus (VSV), we developed a quantitative reverse transcription-PCR assay for the detection, quantification, and differentiation of the major serotypes, VSV Indiana and VSV New Jersey, using a closed-tube multiplex format.
The detection system is based on the recently invented primer-probe energy transfer (PriProET) system. A region of the gene encoding the RNA-dependent RNA polymerase was amplified by using VSV-specific primers in the presence of two serotype-specific fluorescent probes. By incorporating nucleotide analogues in the primers, both serotypes were amplified with similar efficiencies.
The generation of specific amplicons resulted in fluorescent signals for either of the two serotypes, and the specificities of the reactions were confirmed from the melting temperature profiles of the fluorescent probes. The limits of detection ware found to be less than 10 50% tissue culture infective doses/ml for both serotypes.
The diagnostic value of the new method was tested with clinical materials from experimentally infected pigs, and it is concluded that the method is a powerful tool for the rapid identification of VSV.
Language: | English |
---|---|
Publisher: | American Society for Microbiology |
Year: | 2005 |
Pages: | 356-362 |
ISSN: | 1098660x , 00951137 and 1070633x |
Types: | Journal article |
DOI: | 10.1128/JCM.43.1.356-362.2005 |
ORCIDs: | Rasmussen, Thomas Bruun |
Animals Base Sequence DNA Primers Energy Transfer L protein, vesicular stomatitis virus Molecular Sequence Data RNA, Viral RNA-Dependent RNA Polymerase Reverse Transcriptase Polymerase Chain Reaction Rhabdoviridae Infections Stomatitis Swine Swine Diseases Vesicular stomatitis Indiana virus Vesiculovirus Viral Proteins