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Duplex reverse transcription-polymerase chain reaction system to detect Potato spindle tuber viroid using an internal control mRNA and a non-infectious positive control RNA

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Abstract

An accurate and reliable system to detect Potato spindle tuber viroid (PSTVd) was developed based on duplex reverse transcription-polymerase chain reaction (RT-PCR) in combination with a plant mRNA internal control to evaluate the quality of RNA extracted from a dormant potato tuber and the effectiveness of the RT-PCR. In addition, a non-infectious RNA based on the PSTVd sequence was generated in vitro as a readily available positive control without the possibility of escape; this RNA was distinguishable from true positives in PSTVd infection. This detection system will be useful in plant quarantine and phytosanitary certification applications.

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Acknowledgments

This work was supported in part by a grant-in-aid from the Agriculture, Forestry, and Fisheries Research Council, MAFF of Japan (No. 1722).

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  1. Laboratory of Pathogen-Plant Interactions, Research Faculty of Agriculture, Hokkaido University, Kita 9, Nishi 9, Kita-ku, Sapporo, 060-8589, Japan

    Tatsuji Hataya

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  1. Tatsuji Hataya
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Correspondence to Tatsuji Hataya.

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Hataya, T. Duplex reverse transcription-polymerase chain reaction system to detect Potato spindle tuber viroid using an internal control mRNA and a non-infectious positive control RNA. J Gen Plant Pathol 75, 167–172 (2009). https://doi.org/10.1007/s10327-009-0159-z

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  • DOI: https://doi.org/10.1007/s10327-009-0159-z

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