Rapid detection of genetically diverse tomato black ring virus isolates using reverse transcription loop-mediated isothermal amplification

Beata Hasiów-Jaroszewska; Daria Budzyńska; Natasza Borodynko; Henryk Pospieszny

doi:10.1007/s00705-015-2586-9

Rapid detection of genetically diverse tomato black ring virus isolates using reverse transcription loop-mediated isothermal amplification

Arch Virol. 2015 Dec;160(12):3075-8. doi: 10.1007/s00705-015-2586-9. Epub 2015 Sep 4.

Authors

Beata Hasiów-Jaroszewska¹, Daria Budzyńska², Natasza Borodynko², Henryk Pospieszny²

Affiliations

¹ Department of Virology and Bacteriology, Institute of Plant Protection - National Research Institute, ul. Wł. Węgorka 20, 60-318, Poznan, Poland. [email protected].
² Department of Virology and Bacteriology, Institute of Plant Protection - National Research Institute, ul. Wł. Węgorka 20, 60-318, Poznan, Poland.

PMID: 26338092
DOI: 10.1007/s00705-015-2586-9

Abstract

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.

Keywords: Detection; RT-LAMP; TBRV.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

DNA Primers / genetics
Genetic Variation
Nepovirus / classification
Nepovirus / genetics*
Nepovirus / isolation & purification*
Nucleic Acid Amplification Techniques / methods*
Plant Diseases / virology*
Reverse Transcription
Sensitivity and Specificity
Solanum lycopersicum / virology*

Substances

DNA Primers