Development and Verification of Nested PCR Assay for Detection of Tobacco rattle virus in Plant Quarantine.
- Author:
Siwon LEE
1
;
Jin Young LEE
;
Yong Gil SHIN
;
Su Heon LEE
;
Tae Young AHN
Author Information
- Publication Type:Brief Communication
- Keywords: Tobacco rattle virus; Nested PCR; Quarantine
- MeSH: Korea; Lycopersicon esculentum; Plants*; Plasmids; Polymerase Chain Reaction*; Quarantine*; RNA Viruses; Solanum tuberosum; Tobacco*
- From:Journal of Bacteriology and Virology 2015;45(1):54-61
- CountryRepublic of Korea
- Language:Korean
- Abstract: 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.
