Abstract
SYBR Green I based real-time RT-PCR assay was developed for the detection and quantification of duck reovirus (DRV) using ABI PRISM 7500 sequence detection system. The assay was carried out using a set of primer designed to amplify highly conserved sequences of S2 gene of DRV. A 10-fold dilution series assay using a plasmid containing the cDNA of DRV S2 gene demonstrated the high sensitivity of the assay with a lowest detection limit of ≤1.48 copies/μL Standard deviation and coefficient of variation were low for both intra-assay and inter-assay variability. The assay performance was evaluated on 80 samples obtained from artificially infected Cherry Valley ducklings and 10 field specimens compared with the conventional RT-PCR assay. It was shown that 10 artificially infected samples tested negative in gel-based assay were positive for the real-time RT-PCR. DRV could be detected in all eight different tissues collected from the ducklings infected artificially. In contrast, the higher detection rate was obtained in the bursa of fabricius (90%), lung (90%), spleen (80%), and thymus (70%) than that in the liver (30%) as well as in the pancreas (10%). This method was rapid, specific, and sensitive for the detection of DRV and will be useful in veterinary diagnostic applications.
Get full access to this article
View all access options for this article.