title: Development of reverse transcription loop-mediated isothermal amplification assay for rapid detection of an emerging potyvirus: tomato necrotic stunt virus.
abstract: Tomato necrotic stunt virus (ToNStV) is an emerging potyvirus that causes severe stunting to infected tomato plants. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for sensitive detection of ToNStV. The sensitivity of RT-LAMP was comparable to that of conventional RT-PCR, with detection of ToNStV in a reaction containing only 8 pg of total tomato RNA or with 1:20,000 dilution of crude tissue extract. This assay was able to detect ToNStV in a broad range of solanaceous plant species. The RT-LAMP for ToNStV was specific with no cross-reactivity to other potyviruses (i.e. Potato virus Y and Tobacco etch virus), as well as several other common tomato viruses. RT-LAMP should complement RT-PCR and real-time RT-PCR assays reported previously, with a potential to provide a simple, rapid, and sensitive field diagnostic method for ToNStV.
authors: USDA-Agricultural Research Service, U.S. Vegetable Laboratory, Charleston, SC 29414, USA. Electronic address: Rugang.Li@ars.usda.gov.
authors: Rugang
authors: R
authors: Li
authors: USDA-Agricultural Research Service, U.S. Vegetable Laboratory, Charleston, SC 29414, USA. Electronic address: Kai.Ling@ars.usda.gov.
authors: Kai-Shu
authors: KS
authors: Ling
mesh_terms: D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D017800:Potyvirus / Q000235:genetics / Q000302:isolation & purification*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D019657:Solanaceae / Q000821:virology; D014773:Virology / Q000379:methods*
keywords: Field disease diagnosis; LAMP assay; Solanum lycopersicum; qRT-PCR