Rapid detection of two emerging viruses associated with necrotic symptoms in Areca catechu L. by reverse transcription loop-mediated isothermal amplification (RT-LAMP)

J Virol Methods. 2020 Jul:281:113795. doi: 10.1016/j.jviromet.2019.113795. Epub 2019 Dec 3.

Abstract

Two reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed for the detection of areca palm necrotic ringspot virus (ANRSV) and areca palm necrotic spindle-spot virus (ANSSV), respectively. These two emerging viruses both induce necrotic symptoms in areca palms. The coat protein (CP) gene of ANRSV and the 9 K gene of ANSSV were used to design the respective RT-LAMP primers for the assays. Each set of four primers designed for each of these viruses was found to be highly specific in the detection of the respective targeted virus. The optimal incubation conditions for the RT-LAMP assays were 63 °C for 40 min for ANRSV and at 61 °C for 40 min for ANSSV. The sensitivity of the RT-LAMP method for each of these viruses was 10-fold greater than that of the corresponding conventional reverse-transcription polymerase chain reaction (RT-PCR). The RT-LAMP assays may be useful for the rapid early detection of ANSSV and ANRSV in commercial areca palm production.

Keywords: ANRSV; ANSSV; Areca palm; RT-LAMP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Areca / virology*
  • DNA Primers / genetics
  • Molecular Diagnostic Techniques / methods*
  • Nucleic Acid Amplification Techniques / methods*
  • Plant Diseases / virology*
  • RNA, Viral / genetics
  • Reverse Transcription
  • Sensitivity and Specificity
  • Viruses / classification*
  • Viruses / isolation & purification

Substances

  • DNA Primers
  • RNA, Viral

Supplementary concepts

  • LAMP assay