Rapid, specific, simple, in-field detection of Xanthomonas campestris pathovar musacearum by loop-mediated isothermal amplification
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Hodgetts, J.; Hall, J.; Karamura, G.; Grant, M.; Studholme, D.J.; Boonham, N.; Karamura, E.; Smith, J.J. (2015) Rapid, specific, simple, in-field detection of Xanthomonas campestris pathovar musacearum by loop-mediated isothermal amplification. Journal of Applied Microbiology 119 (6) p. 1651–1658 ISSN: 1364-5072
Permanent link to this item: http://hdl.handle.net/10568/69063
Internet URL: http://onlinelibrary.wiley.com/doi/10.1111/jam.12959/abstract;jsessionid=181C335463C8A8DCCB17985ED0A9EDD8.f01t04?systemMessage=Wiley+Online+Library+will+have+be+unavailable+on+Saturday+5th+December+from+10%3A00-14%3A00+GMT+%2F+05%3A00-09%3A00+EST+%2F+18%3A00-22%3A00+SGT+for+essential+maintenance.+Apologies+for+the+inconvenience.
The aim of this research was 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. 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. 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. This is the first LAMP assay for Xcm which provides a significant improvement compared to existing diagnostics.