LNA probe-based assay for the detection of Tomato black ring virus isolates

Mol Cell Probes. 2015 Feb;29(1):78-80. doi: 10.1016/j.mcp.2014.12.002. Epub 2014 Dec 24.

Abstract

Tomato black ring virus (TBRV) infects a wide range of economically important plant species worldwide. In the present study we developed a locked nucleic acid (LNA) real-time RT-PCR assay for accurate detection of genetically diverse TBRV isolates collected from different hosts. The assay based on the LNA probe has a wide detection range, high sensitivity, stability and amplification efficiency. The assay amplified all tested TBRV isolates, but no signal was observed for the RNA from other nepoviruses and healthy plant species. Under optimum reaction conditions, the detection limit was estimated around 17 copies of the TBRV target region in total RNA. Real-time RT-PCR with the LNA probe described in this paper will serve as a valuable tool for robust, sensitive and reliable detection of TBRV isolates.

Keywords: Detection; LNA probe; Real-time RT-PCR; Tomato black ring virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Nepovirus / classification*
  • Nepovirus / isolation & purification*
  • Oligonucleotide Probes / genetics*
  • Plant Diseases / virology
  • RNA, Viral / analysis
  • Real-Time Polymerase Chain Reaction / methods
  • Sensitivity and Specificity
  • Solanum lycopersicum / virology*

Substances

  • Oligonucleotide Probes
  • RNA, Viral