title: Loop-mediated isothermal amplification for the detection of plant pathogens.
abstract: Loop-mediated isothermal amplification (LAMP) is a technique involving the use of four to six primers (two inner primers, two outer primers, and two loop primers) and the strand displacement activity of Bacillus subtilis-derived (Bst) DNA polymerase. The end result of strand displacement and loop formation and synthesis is the single-temperature amplification of a highly specific fragment from a DNA template at a much greater titre than that obtained with polymerase chain reaction. With LAMP, there are several methods to determine a positive reaction. Presented here are three alternative methods: gel electrophoresis, hydroxynaphthol blue colorimetric dye, and the fluorescent intercalating PicoGreen(®) reagent.
authors.affiliation: Plant Health and Environment Laboratory, Investigation and Diagnostic Centre, MAF Biosecurity New Zealand, Wellington, New Zealand. lisa.ward@maf.govt.nz
authors.forename: Lisa I
authors.initials: LI
authors.lastname: Ward
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Harper
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D010944:Plants / Q000235:genetics / Q000382:microbiology*; D044167:Xylella / Q000235:genetics
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D004259:DNA-Directed DNA Polymerase