Journal article
Development of a primer-probe energy transfer based real-time PCR for detection of Marek's disease virus
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 TCID50/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.
Language: | English |
---|---|
Year: | 2010 |
Pages: | 21-26 |
ISSN: | 18790984 and 01660934 |
Types: | Journal article |
DOI: | 10.1016/j.jviromet.2009.12.012 |
ORCIDs: | Rasmussen, Thomas Bruun |