Development of a primer-probe energy transfer based real-time PCR for detection of Marek's disease virus
| Autor: | Peer Lyng Frandsen, Erik Østergaard, Annette Malene Barfoed, Thomas Bruun Rasmussen, Eva Sandberg, Eva Bækdahl Nielsen |
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| Rok vydání: | 2009 |
| Předmět: |
Molecular Sequence Data
Biology Polymerase Chain Reaction Sensitivity and Specificity Virus law.invention law Virology Marek Disease Animals Transition Temperature Polymerase chain reaction Marek's disease Base Sequence Amplicon biology.organism_classification Molecular biology Virus Cultivation Mardivirus Real-time polymerase chain reaction Capsid Energy Transfer Capsid Proteins Primer (molecular biology) Oligonucleotide Probes Sequence Alignment |
| Zdroj: | Journal of virological methods. 165(1) |
| ISSN: | 1879-0984 |
| Popis: | A real-time PCR assay, which enables simultaneous detection and differentiation of all three serotypes of Marek's disease virus, without the need for post-PCR sequencing, has been developed. The assay is based on the primer-probe energy transfer real-time PCR, which has a relatively high tolerance towards point mutations in the probe region. The PCR is followed by a probe melting point analysis, which enables confirmation of identity of amplicon and differentiation of serotypes. The assay targets the MDV031 gene, encoding UL19 major capsid protein-like protein and was shown to be quantitative, with a detection limit below 10 TCID 50 /ml starting material. This sensitivity is similar to the one obtained with traditional virus cultivation. However, the PCR method can provide a laboratory result within a day, while the virus cultivation method takes more than a week to perform. The new method will be useful for testing of avian live viral vaccines and screening for extraneous agents. |
| Databáze: | OpenAIRE |
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