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

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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.

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Acknowledgments

This work was partially supported by National Science Centre project 2013/11/B/NZ9/02510 and Iuventus Plus IP2014 014973 (years 2015–2017) from the Ministry of Science and Higher Education.

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Authors and Affiliations

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

    Beata Hasiów-Jaroszewska, Daria Budzyńska, Natasza Borodynko & Henryk Pospieszny

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  1. Beata Hasiów-Jaroszewska
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  2. Daria Budzyńska
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  3. Natasza Borodynko
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  4. Henryk Pospieszny
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Corresponding author

Correspondence to Beata Hasiów-Jaroszewska.

Electronic supplementary material

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705_2015_2586_MOESM1_ESM.tif

Supplementary file Alignment of TBRV sequences used for designing the primers and their location within the sequences (TIFF 2023 kb)

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Hasiów-Jaroszewska, B., Budzyńska, D., Borodynko, N. et al. Rapid detection of genetically diverse tomato black ring virus isolates using reverse transcription loop-mediated isothermal amplification. Arch Virol 160, 3075–3078 (2015). https://doi.org/10.1007/s00705-015-2586-9

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  • DOI: https://doi.org/10.1007/s00705-015-2586-9

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