Real-time loop-mediated isothermal amplification assay for rapid detection of Fusarium circinatum

Biotechniques. 2020 Jul;69(1):369-375. doi: 10.2144/btn-2019-0168. Epub 2020 Apr 27.

Abstract

Fusarium circinatum is the causal agent of pitch canker, a lethal disease of pine and other conifers. Since F. circinatum is a quarantine organism, its timely detection could efficiently prevent its introduction into new areas or facilitate spread management in already infected sites. In this study, we developed a sequence-specific probe loop-mediated isothermal amplification (LAMP) assay for F. circinatum using a field-deployable portable instrument. The assay was able to recognize the pathogen in host tissues in just 30 min, and the sensitivity of the assay made it possible to detect even small amounts of F. circinatum DNA (as low as 0.5 pg/μl). The high efficiency of this method suggests its use as a standard diagnostic tool during phytosanitary controls.

Keywords: LAMP; elongation factor gene; field-deployable; invasive species; isothermal amplification; pine pitch canker.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Fungal / genetics
  • Fusarium / genetics*
  • Molecular Diagnostic Techniques / methods*
  • Nucleic Acid Amplification Techniques / methods*
  • Plant Diseases / microbiology
  • Sensitivity and Specificity

Substances

  • DNA, Fungal

Supplementary concepts

  • Fusarium circinatum
  • LAMP assay