title: Rapid detection of tobacco viruses by reverse transcription loop-mediated isothermal amplification.
abstract: Tobacco viruses may cause a wide range of diseases that heavily reduce tobacco quality and yield worldwide. In order to detect viral diseases in tobacco fields, a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) method was established. Nucleotide amplification could be observed clearly after adding SYBR Green I, within 60 min under isothermal conditions, at 63-65 °C with a set of primers targeting the viral coat protein (CP) genes of tobacco viruses including cucumber mosaic virus (CMV), potato virus Y (PVY), tobacco etch virus (TEV), tobacco mosaic virus (TMV) and tobacco vein banding mosaic virus (TVBMV). This method has high specificity and sensitivity. The sensitivity of the RT-LAMP was 10 to 100 times higher than that of the conventional RT-PCR method. The RT-LAMP assay was proven reliable for virus diagnosis of tobacco samples from the field.
authors.affiliation: Key Laboratory of Crop Pest Integrated Pest Management on Crop in Northwestern Loess Plateau, Ministry of Agriculture, Key Laboratory of Plant Protection Resources and Pest Management, Ministry of Education, State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling 712100, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhao
authors.forename: Julong
authors.initials: J
authors.lastname: Cheng
authors.forename: Xingan
authors.initials: X
authors.lastname: Hao
authors.forename: Xiaoman
authors.initials: X
authors.lastname: Tian
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Wu
mesh_terms: D002681:China; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000191:economics / Q000821:virology; D010942:Plant Viruses / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D014026:Nicotiana / Q000821:virology*
chemical_list: D012367:RNA, Viral