Comparison of DAS-ELISA and RT-PCR Methods for the Detection of Prunus Necrotic Ringspot Virus (PNRSV)
نویسندگان
چکیده
Two methods (DAS-ELISA and RT-PCR) were compared for the detection of Prunus necrotic ringspot virus (PNRSV) in woody host Prunus mahleb isolated from Malatya. Total RNA extractions were made from serially diluted fresh leaf tissue, root, bark and one year old green bark tissue using silica-based method. Purified total RNA extracts were used as template for cDNA synthesis by reverse transcription. PCR products, about 616 base pair in size, was analyzed by gel electrophoresis and visualized by ethidium bromide staining. Comparing ELISA test, RT-PCR was found more sensitive in detecting local PNRSV isolate. Several plant tissues were used for the evaluation of best tissue in the detection of PNRSV by RT-PCR. Leaf tissue of infected P. mahaleb was proved to be more suitable tissue in detection of virus by RT-PCR respect to root, one-year-old green bark tissue and bark tissue.
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