Loop-Mediated Isothermal Amplification (LAMP) for Detection of Phytoplasmas in the Field

Methods Mol Biol. 2015:1302:99-111. doi: 10.1007/978-1-4939-2620-6_8.

Abstract

Loop-mediated isothermal amplification (LAMP) is a nucleic acid-based detection method with many applications. This chapter details its use for detection of phytoplasma diseases, combining a rapid 2-min DNA extraction method with real-time LAMP product detection such that the entire procedure can be undertaken and completed in the field within 40 min. Furthermore, the assays include an anneal curve validation step to guard against false positives and an assay for host plant DNA to guard against false negatives.

MeSH terms

  • Catharanthus / genetics
  • Catharanthus / microbiology*
  • DNA, Bacterial / analysis*
  • DNA, Bacterial / genetics
  • DNA, Plant / analysis*
  • DNA, Plant / genetics
  • Nucleic Acid Amplification Techniques / methods*
  • Phytoplasma / genetics
  • Phytoplasma / isolation & purification*
  • Phytoplasma / pathogenicity
  • Plant Diseases / microbiology*
  • Plant Leaves / genetics
  • Plant Leaves / microbiology

Substances

  • DNA, Bacterial
  • DNA, Plant