BioTechniques (Jul 2020)

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

  • Dagmar Stehlíková,
  • Nicola Luchi,
  • Chiara Aglietti,
  • Alessia Lucia Pepori,
  • Julio Javier Diez,
  • Alberto Santini

DOI
https://doi.org/10.2144/btn-2019-0168
Journal volume & issue
Vol. 69, no. 1
pp. 11 – 17

Abstract

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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.

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