Abstract
A one-step multiplex quantitative reverse transcription polymerase chain reaction protocol is described, for the detection in pome trees of Pear blister canker viroid and Apple scar skin viroid, together with universal detection of phytoplasmas. Total nucleic acids extraction is performed according to a modified CTAB protocol and TaqMan MGB probes are used to surpass high genetic variability of viroids. The multiplex real-time assay is at least ten times more sensitive than conventional protocols and its features make it suitable for rapid and massive screening of pome fruit trees phytoplasmas and viroids in certification schemes and surveys.
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Acknowledgment
This work was supported by the Hellenic GSRT Project “09SYN-22-638” and partially by FP7- REGPOT-2008-1 project “BPI-PlantHeal 230010.”
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Malandraki, I., Varveri, C., Vassilakos, N. (2019). One-Step Multiplex Quantitative RT-PCR for the Simultaneous Detection of Viroids and Phytoplasmas. In: Musetti, R., Pagliari, L. (eds) Phytoplasmas. Methods in Molecular Biology, vol 1875. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8837-2_12
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DOI: https://doi.org/10.1007/978-1-4939-8837-2_12
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-8837-2
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