title: Development of multiplex real-time PCR for simultaneous detection of three Potyviruses in tobacco plants.
abstract: OBJECTIVE
  To develop a multiplex real-time PCR assay using TaqMan probes for the simultaneous detection and quantification of Tobacco etch virus (TEV), Potato virus Y (PVY) and Tobacco vein banding mosaic virus (TVBMV).
  
  
  RESULTS
  Specific primer and probe combinations for TEV and TVBMV were developed from the coat protein region of the viral genome. To detect PVY, a primer and probe combination PVY-Univ F, PVY-Univ R and PVY-Univ P for amplifying the coat protein region of the virus genome was employed. The detection limit of multiplex real-time PCR for these viruses was 10 copies μl(-1) of the standard plasmid. The multiplex reaction was successful in the detection of these three pathogens, with no non-specific amplification and cross-reaction.
  
  
  CONCLUSIONS
  This multiplex real-time PCR provides a rapid, effective, specific and sensitive method for the simultaneous detection and quantification of the three pathogens on infected tobacco plants.
  
  
  CONCLUSIONS
  This multiplex real-time PCR will be useful not only for diagnostic, ecological, epidemiological and pathogenesis studies, but also for investigating host/virus or virus/virus interactions, in particular during mix infection.
authors.affiliation: State Key Laboratory of Crop Stress Biology in Arid Areas and Key Laboratory of Crop Pest Integrated Pest Management on the Loess Plateau of Ministry of Agriculture, College of Plant Protection, Northwest A&F University, Yangling, China.
authors.forename: J
authors.initials: J
authors.lastname: Dai
authors.forename: H
authors.initials: H
authors.lastname: Peng
authors.forename: W
authors.initials: W
authors.lastname: Chen
authors.forename: J
authors.initials: J
authors.lastname: Cheng
authors.forename: Y
authors.initials: Y
authors.lastname: Wu
mesh_terms: D017931:DNA Primers; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D015341:Nucleic Acid Probes; D017800:Potyvirus / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014026:Nicotiana / Q000821:virology*
chemical_list: D017931:DNA Primers; D015341:Nucleic Acid Probes