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Genome-informed diagnostics for specific and rapid detection of Pectobacterium species using recombinase polymerase amplification coupled with a lateral flow device
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Genome-informed diagnostics for specific and rapid detection of Pectobacterium species using recombinase polymerase amplification coupled with a lateral flow device
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Journal Article
Genome-informed diagnostics for specific and rapid detection of Pectobacterium species using recombinase polymerase amplification coupled with a lateral flow device
2018
Overview
Pectobacterium
species cause serious bacterial soft rot diseases worldwide on economically important fruit and vegetable crops including tomato and potato. Accurate and simple methods are essential for rapid pathogen identification and timely management of the diseases. Recombinase polymerase amplification (RPA) combined with a lateral flow device (LFD) was developed for specific detection of
Pectobacterium
sp. directly from infected plant materials with no need for DNA isolation. The specificity of RPA-LFD was tested with 26
Pectobacterium
sp. strains and 12 non-
Pectobacterium
species and no false positive or false negative outcomes were observed. RPA primers and probe for host control were also developed to detect the host genome for enhanced reliability and accuracy of the developed assay. The detection limit of 10 fg was obtained with both sensitivity and spiked sensitivity assays. No inhibitory effects were observed on the RPA assay when targets (pathogen and host) were directly detected from infected potato and tomato sap. The developed RPA assay has numerous applications from routine diagnostics at point-of-care, biosecurity, surveillance and disease management to epidemiological studies. In addition, this tool can also be used to discover reservoir hosts for
Pectobacterium
species.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 45/23
/ 45/29
/ 45/43
/ 45/77
/ Bacterial Proteins - genetics
/ DNA
/ DNA-Binding Proteins - genetics
/ Genomes
/ Humanities and Social Sciences
/ Loop-mediated Isothermal Amplification (LAMP)
/ Nucleic Acid Amplification Techniques - instrumentation
/ Nucleic Acid Amplification Techniques - methods
/ Pectobacterium - classification
/ Pectobacterium - isolation & purification
/ Potatoes
/ Primers
/ Recombinase Polymerase Amplification (RPA)
/ Science
/ Soft rot
/ Species
/ Tomatoes
