title: Rapid, specific, simple, in-field detection of Xanthomonas campestris pathovar musacearum by loop-mediated isothermal amplification.
abstract: OBJECTIVE
  To develop and evaluate a loop-mediated isothermal amplification (LAMP) assay for Xanthomonas campestris pathovar musacearum (Xcm), the causal agent of banana Xanthomonas wilt, a major disease of banana in Africa.
  
  
  RESULTS
  LAMP primers were designed to the general secretion pathway protein D gene and tested against 17 isolates of Xcm encompassing the known genetic and geographic diversity of the bacterium and all isolates were detected. Seventeen other Xanthomonas isolates, including closely related Xanthomonas vasicola, other bacterial pathogens/endophytes of Musa and two healthy Musa varieties gave negative results with the LAMP assay. The assay showed good sensitivity, detecting as little as 51 fg of Xcm DNA, a greater level of sensitivity than that of an Xcm PCR assay. Amplification with the LAMP assay was very rapid, typically within 9 min from bacterial cultures. Symptomatic field samples of Musa from Uganda were tested and all produced amplification in less than 13 min.
  
  
  CONCLUSIONS
  The LAMP assay provides rapid, sensitive detection of the pathogen that is ideally suited for deployment in laboratories with basic facilities and in-field situations.
  
  
  CONCLUSIONS
  This is the first LAMP assay for Xcm which provides a significant improvement compared to existing diagnostics.
authors.affiliation: Fera, National Agri-Food Innovation Campus, York, UK.
authors.forename: J
authors.initials: J
authors.lastname: Hodgetts
authors.affiliation: Fera, National Agri-Food Innovation Campus, York, UK.
authors.forename: J
authors.initials: J
authors.lastname: Hall
authors.affiliation: Fera, National Agri-Food Innovation Campus, York, UK.|Biosciences, College of Life and Environmental Sciences, University of Exeter, Exeter, Devon, UK.
authors.forename: G
authors.initials: G
authors.lastname: Karamura
authors.affiliation: Biosciences, College of Life and Environmental Sciences, University of Exeter, Exeter, Devon, UK.
authors.forename: M
authors.initials: M
authors.lastname: Grant
authors.affiliation: Biosciences, College of Life and Environmental Sciences, University of Exeter, Exeter, Devon, UK.
authors.forename: D J
authors.initials: DJ
authors.lastname: Studholme
authors.affiliation: Fera, National Agri-Food Innovation Campus, York, UK.
authors.forename: N
authors.initials: N
authors.lastname: Boonham
authors.affiliation: Bioversity International/CRP-RTB, Nairobi, Kenya.
authors.forename: E
authors.initials: E
authors.lastname: Karamura
authors.affiliation: Fera, National Agri-Food Innovation Campus, York, UK.
authors.forename: J J
authors.initials: JJ
authors.lastname: Smith
mesh_terms: D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D028521:Musa / Q000382:microbiology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000382:microbiology*; D016959:Xanthomonas campestris / Q000235:genetics / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial
keywords: Xanthomonas campestris pathovar musacearum; bacterial phytopathogens; banana Xanthomonas wilt; diagnostics; in-field testing; loop-mediated isothermal amplification; rapid detection