Erwinia amylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight

Bühlmann, Andreas; Pothier, F.; Rezzonico, Fabio; Smits, Theo H. M.; Andreou, Michael; Boonham, Neil; Duffy, Brion; Frey, Jürg E.

doi:10.1016/j.mimet.2012.12.017

Publication type:	Article in scientific journal
Type of review:	Peer review (publication)
Title:	Erwinia amylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight
Authors:	Bühlmann, Andreas Pothier, F. Rezzonico, Fabio Smits, Theo H. M. Andreou, Michael Boonham, Neil Duffy, Brion Frey, Jürg E.
DOI:	10.1016/j.mimet.2012.12.017
Published in:	Journal of Microbiological Methods
Volume(Issue):	92
Issue:	3
Page(s):	332
Pages to:	339
Issue Date:	Mar-2013
Publisher / Ed. Institution:	Elsevier
ISSN:	0167-7012
Language:	English
Subject (DDC):	570: Biology 630: Agriculture
Abstract:	Several molecular methods have been developed for the detection of Erwinia amylovora, the causal agent of fire blight in pear and apple, but none are truly applicable for on-site use in the field. We developed a fast, reliable and field applicable detection method using a novel target on the E. amylovora chromosome that we identified by applying a comparative genomic pipeline. The target coding sequences (CDSs) are both uniquely specific for and all-inclusive of E. amylovora genotypes. This avoids potential false negatives that can occur with most commonly used methods based on amplification of plasmid gene targets, which can vary among strains. Loop-mediated isothermal AMPlification (LAMP) with OptiGene Genie II chemistry and instrumentation proved to be an exceptionally rapid (under 15 min) and robust method for detecting E. amylovora in orchards, as well as simple to use in the plant diagnostic laboratory. Comparative validation results using plant samples from inoculated greenhouse trials and from natural field infections (of regional and temporal diverse origin) showed that our LAMP had an equivalent or greater performance regarding sensitivity, specificity, speed and simplicity than real-time PCR (TaqMan), other LAMP assays, immunoassays and plating, demonstrating its utility for routine testing.
URI:	https://digitalcollection.zhaw.ch/handle/11475/1727
Fulltext version:	Published version
License (according to publishing contract):	Licence according to publishing contract
Departement:	Life Sciences and Facility Management
Organisational Unit:	Institute of Natural Resource Sciences (IUNR)
Appears in collections:	Publikationen Life Sciences und Facility Management

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Bühlmann, A., Pothier, F., Rezzonico, F., Smits, T. H. M., Andreou, M., Boonham, N., Duffy, B., & Frey, J. E. (2013). Erwinia amylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight. Journal of Microbiological Methods, 92(3), 332–339. https://doi.org/10.1016/j.mimet.2012.12.017

Bühlmann, A. et al. (2013) ‘Erwinia amylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight’, Journal of Microbiological Methods, 92(3), pp. 332–339. Available at: https://doi.org/10.1016/j.mimet.2012.12.017.

A. Bühlmann et al., “Erwinia amylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight,” Journal of Microbiological Methods, vol. 92, no. 3, pp. 332–339, Mar. 2013, doi: 10.1016/j.mimet.2012.12.017.

BÜHLMANN, Andreas, F. POTHIER, Fabio REZZONICO, Theo H. M. SMITS, Michael ANDREOU, Neil BOONHAM, Brion DUFFY und Jürg E. FREY, 2013. Erwinia amylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight. Journal of Microbiological Methods. März 2013. Bd. 92, Nr. 3, S. 332–339. DOI 10.1016/j.mimet.2012.12.017

Bühlmann, Andreas, F. Pothier, Fabio Rezzonico, Theo H. M. Smits, Michael Andreou, Neil Boonham, Brion Duffy, and Jürg E. Frey. 2013. “Erwinia Amylovora Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Pathogen Detection and On-Site Diagnosis of Fire Blight.” Journal of Microbiological Methods 92 (3): 332–39. https://doi.org/10.1016/j.mimet.2012.12.017.

Bühlmann, Andreas, et al. “Erwinia Amylovora Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Pathogen Detection and On-Site Diagnosis of Fire Blight.” Journal of Microbiological Methods, vol. 92, no. 3, Mar. 2013, pp. 332–39, https://doi.org/10.1016/j.mimet.2012.12.017.