Rapid and Sensitive Detection of Citrus tristeza virus Using Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Assay

  • Dilip Kumar Ghosh
  • Ashish Warghane
  • Kajal Kumar Biswas
Part of the Methods in Molecular Biology book series (MIMB, volume 2015)


Loop-mediated isothermal amplification (LAMP) is one recently developed gene amplification technique that emerges as a simple and quick diagnostic tool for early detection of nucleic acid targets. The LAMP technique works on the principle of strand displacement activity of Bst polymerase. It contains a set of four specially designed primers, which recognizes six different regions on the target nucleotide sequence. In the LAMP reaction, amplification is carried out in an isothermal conditions (60–65°C) using simple and inexpensive device like water bath or dry bath. Additional benefits of LAMP technique are that final results can be seen directly with naked eyes by adding intercalating dye SYBR Green I in the reaction tube. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is one of the novel techniques used for detection of RNA targets. The technology has been successfully applied for rapid and sensitive detection of Citrus tristeza virus (CTV) by using four oligo-primers, targeting a conserved coat protein gene (CPG) of an Indian CTV isolate. The result of assay is visible in naked eyes easily in the presence of SYBR Green I (100×) or on 1.5% agarose gel electrophoresis. CTV-RT-LAMP could be used away from plant pathology laboratories even in remote location.

Key words

Citrus Tristeza disease RT-LAMP SYBR Green I Bst DNA polymerase 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Dilip Kumar Ghosh
    • 1
  • Ashish Warghane
    • 1
  • Kajal Kumar Biswas
    • 2
  1. 1.Plant Virology LaboratoryICAR-Central Citrus Research InstituteNagpurIndia
  2. 2.Division of Plant Pathology, Advanced Centre for Plant VirologyICAR Indian Agricultural Research InstituteNew DelhiIndia

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