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Rapid, specific, simple, in-field detection ofXanthomonas campestrispathovarmusacearumby loop-mediated isothermal amplification

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Hodgetts, J., Hall, J., Karamura, G., Grant, Murray, Studholme, David J., Boonham, N., Karamura, E. and Smith, J. J. (2015) Rapid, specific, simple, in-field detection ofXanthomonas campestrispathovarmusacearumby loop-mediated isothermal amplification. Journal of Applied Microbiology, 119 (6). pp. 1651-1658. doi:10.1111/jam.12959 ISSN 1364-5072.

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Official URL: http://dx.doi.org/10.1111/jam.12959

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Abstract

Aims

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.
Methods and 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.

Item Type: Journal Article
Divisions: Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- )
Journal or Publication Title: Journal of Applied Microbiology
Publisher: Wiley-Blackwell Publishing Ltd.
ISSN: 1364-5072
Official Date: November 2015
Dates:
DateEvent
November 2015Published
22 November 2015Available
1 October 2015Accepted
19 July 2015Submitted
Volume: 119
Number: 6
Page Range: pp. 1651-1658
DOI: 10.1111/jam.12959
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Restricted or Subscription Access
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