期刊
JOURNAL OF VIROLOGICAL METHODS
卷 246, 期 -, 页码 90-94出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2017.04.014
关键词
Foot-and-mouth disease virus; Real-time RT-PCR; 5 '-tailed primers; Validation
资金
- CODA-CERVA
- Belgian Federal Public Service for Health, Food Chain Safety and Environment
- Belgian Federal Agency for the Safety of the Food Chain (FAVVAFSCA)
The 3D and 5UTR real-time RT-PCR assays (RT-qPCR) from Callahan et al. (2002) and Reid et al. (2002) are commonly used reference methods for the detection of foot-and-mouth disease virus (FMDV). For an optimal detection of FMDV in clinical samples, it is advised to use both assays simultaneously (King et al., 2006). Recently, Vandenbussche et al. (2016) showed that the addition of 5 '-tails to the FMDV-specific primers enhances the detection of FMDV in both the 3D and the 5UTR RT-qPCR assay. To validate the 3D and 5UTR RT-qPCR assays with 5 '-tailed primers for diagnostic purposes, both assays were run in parallel in a triplex one-step RT-qPCR protocol with beta-actin as an internal control and synthetic RNA as an external control. We obtained low limits of detection and high linearity's, high repeatability and reproducibility, near 100% analytical specificity and > 99% diagnostic accuracy for both assays. It was concluded that the 3D and 5UTR RT-qPCR assays with 5 '-tailed primers are particularly suited for the detection of FMDV as well as to exclude the presence of FMDV.
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