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Rapid, specific, simple, in-field detection of Xanthomonas campestris pathovar musacearum by loop-mediated isothermal amplification

Lookup NU author(s): Professor Neil Boonham

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Abstract

© 2015. The Society for Applied Microbiology.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. Significance and Impact of the Study: This is the first LAMP assay for Xcm which provides a significant improvement compared to existing diagnostics.


Publication metadata

Author(s): Hodgetts J, Hall J, Karamura G, Grant M, Studholme DJ, Boonham N, Karamura E, Smith JJ

Publication type: Article

Publication status: Published

Journal: Journal of Applied Microbiology

Year: 2015

Volume: 119

Issue: 6

Pages: 1651-1658

Print publication date: 01/12/2015

Online publication date: 01/10/2015

Acceptance date: 21/09/2015

ISSN (print): 1364-5072

ISSN (electronic): 1365-2672

Publisher: Wiley-Blackwell

URL: https://doi.org/10.1111/jam.12959

DOI: 10.1111/jam.12959

PubMed id: 26425811


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