J Bacteriol Virol.  2015 Mar;45(1):54-61. 10.4167/jbv.2015.45.1.54.

Development and Verification of Nested PCR Assay for Detection of Tobacco rattle virus in Plant Quarantine

Affiliations
  • 1Environmental Infrastructure Research Department, National Institute of Environmental Research, Incheon, Korea.
  • 2Incheon International Airport Regional Office, Animal and Plant Quarantine Agency, Incheon, Korea.
  • 3School of Applied Biosciences, Kyungpook National University, Daegu, Korea. suheon@knu.ac.kr
  • 4Department of Microbiology, College of Natural Sciences, Dankook University, Cheonan, Korea. ahnty@dankook.ac.kr

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.

Keyword

Tobacco rattle virus; Nested PCR; Quarantine

MeSH Terms

Korea
Lycopersicon esculentum
Plants*
Plasmids
Polymerase Chain Reaction*
Quarantine*
RNA Viruses
Solanum tuberosum
Tobacco*
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