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KMID : 0921620150450010054
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Journal of Bacteriology and Virology
2015 Volume.45 No. 1 p.54 ~ p.61
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Development and Verification of Nested PCR Assay for Detection of Tobacco rattle virus in Plant Quarantine
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Abstract
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Tobacco rattle virus (TRV) is a plant pathogen belonging to the Group IV positive-sense single-stranded RNA viruses. TRV causes disease in various plants (e.g., potato, tomato and tobacco), for which it was classified as a controlled quarantine virus in Korea. This study aimed to develop specific primer sets for the rapid detection of TRV. Two RT-PCR primer sets were developed for specific detection of TRV. Furthermore, nested primer sets were also developed, which is required for high sensitivity detection in plant quarantine. The RT-PCR and nested PCR products had the following sizes: set 5 (1,096→540 bp), and set 7 (878→756 bp), respectively. In addition, a modified positive-control plasmid was also developed for use as a positive control in TRV quarantine. The diagnostic system for TRV detection was verified using samples from Korean quarantine sites for the last five years (2009-2014). A total of 83 cases were detected among various import crops. This system for detection of TRV will continuously contribute to plant quarantine in the future.
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KEYWORD
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Tobacco rattle virus, Nested PCR, Quarantine
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