@article { author = {Rao, Xueqin and Sun, Jie}, title = {Development of SYBR Green I Based Real-Time RT-PCR Assay for Specific Detection of Watermelon silver mottle Virus}, journal = {Iranian Journal of Biotechnology}, volume = {13}, number = {3}, pages = {20-24}, year = {2015}, publisher = {National Institute of Genetic Engineering and Biotechnology of Iran}, issn = {1728-3043}, eissn = {2322-2921}, doi = {10.15171/ijb.1124}, abstract = {Background: Watermelon silver mottle virus (WSMoV), which belongs to the genus Tospovirus, causes significant loss in Cucurbitaceae plants. Objectives: Development of a highly sensitive and reliable detection method for WSMoV. Materials and Methods: Recombinant plasmids for targeting the sequence of nucleocapsid protein gene of WSMoV were constructed. SYBR Green I real-time PCR was established and evaluated with standard recombinant plasmids and 27 watermelon samples showing WSMoV infection symptoms. Results: The recombinant plasmid was used as template for SYBR Green I real-time PCR to generate standard and melting curves. Melting curve analysis indicated no primer-dimers and non-specific products in the assay. No cross-reaction was observed with Capsicum chlorosis virus (genus Tospovirus) and Cucumber mosaic virus (genus Cucumovirus). Repeatability tests indicated that inter-assay variability of the Ct values was 1.6%. Conclusions: A highly sensitive, reliable and rapid detection method of SYBR Green I real-time PCR for timely detection of WSMoV plants and vector thrips was established, which will facilitate disease forecast and control.}, keywords = {Real-Time RT-PCR,SYBR Green I,Watermelon silver mottle virus}, url = {https://www.ijbiotech.com/article_11625.html}, eprint = {https://www.ijbiotech.com/article_11625_e2b64d8d9c3cf66a43ef7c3b12a99388.pdf} }