Rapid detection of genetically diverse tomato black ring virus isolates using reverse transcription loop-mediated isothermal amplification.
A reverse transcription loop-mediated isothermal amplification assay (RT-LAMP) has been developed for detection of tomato black ring virus (TBRV) isolates collected from different hosts. One-step RT-LAMP was performed with a set of four primers, the design of which was based on the coat protein gene. Results of RT-LAMP were visualized by direct staining of products with fluorescent dyes, agarose gel electrophoresis, and analysis of amplification curves. The sensitivity of RT-LAMP was 100-fold greater than that of RT-PCR. The RT-LAMP assay developed here is a useful and practical method for diagnosis of TBRV.
Department of Virology and Bacteriology, Institute of Plant Protection - National Research Institute, ul. Wł. Węgorka 20, 60-318, Poznan, Poland. beatahasiow@tlen.pl.
Beata
B
Hasiów-Jaroszewska
Department of Virology and Bacteriology, Institute of Plant Protection - National Research Institute, ul. Wł. Węgorka 20, 60-318, Poznan, Poland.
Daria
D
Budzyńska
Department of Virology and Bacteriology, Institute of Plant Protection - National Research Institute, ul. Wł. Węgorka 20, 60-318, Poznan, Poland.
Natasza
N
Borodynko
Department of Virology and Bacteriology, Institute of Plant Protection - National Research Institute, ul. Wł. Węgorka 20, 60-318, Poznan, Poland.
Henryk
H
Pospieszny
D017931:DNA Primers / Q000235:genetics; D014644:Genetic Variation; D018551:Solanum lycopersicum / Q000821:virology*; D017860:Nepovirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity
D017931:DNA Primers
Detection; RT-LAMP; TBRV