Abstract
Pectobacterium atrosepticum (Pba) is a quarantine and threatening phytopathogen known as the causal agent of blackleg and soft rot disease of potatoes in many areas. Its early detection is then important to have healthy potato tubers and reduce economic losses. Today, conventional methods such as enzyme-linked immunosorbent-assay (ELISA) and polymerase chain reaction (PCR) are typically used for Pba detection, but they are expensive and time-consuming. Here we report on the optimization of an alternative approach based on an electrochemical impedance immunosensor combining a microfluidic module and a microelectrodes array, and having advantages in terms of low cost, ease of use and portability. For validation and for assessing its performance, the lab-on-chip platform has been compared with two standard methods (ELISA and PCR).
Highlights
Two other bacteria sources belonging to other Pectobacreium and Dickeya species (Pectobacterium parmentieri strain Wpp161, Dickeya solani strain 2222) were obtained from the collection of the James Hutton Institute (Scotland, United Kingdom) and used as a negative control for diagnostic specificity
enzyme-linked immunosorbent-assay (ELISA) cross-reaction was not observed against the plant pathogens: Agrobacterium tumefaciens; Erwinia amylovora, Pseudomonas marginalis, Ralstonia solanacearum; Xanthomonas campestris, Klebsiella oxycota, Escherichia coli, Rhodococcus rhodochrous, Curtobacterium flaccumfaciens, Pectobacterium parmentieri, P. carotovorum sub sp carotovourum, P. carotovorum sub sp brasiliense, Dickeya
Two 690 bp and 439 bp DNA fragments were respectively produced for P. atrosepticum isolates, while no amplification was observed with isolates of other Pectobacterium spp. and Dickeya spp
Summary
The agrifood field recently exhibited a strong need for new tools to fight pathogen infections by bacteria, fungi and/or viruses, which every year cause worldwide 20% crop losses and 10% post-harvest losses. The bacterium may be concentrated in potato stems and tubers and live there for a significant time in latent form without showing symptoms, but P. atrosepticum can produce huge amounts of extracellular pectinolytic enzymes which leads to specific decay symptoms. This can result in the destruction of the plant cell wall and the middle lamella of leaves, stems, roots, and tubers (Figure 1). As there are not sufficiently effective protocols for mass treatment, one of the best agro-technical approaches is to reject the infected plants For all these reasons, timely detecting P. atrosepticum is crucial to effectively protect plants and produce potato seed tubers [7,8]. In order to monitor Pba in the field, it would be desirable to have access to faster, simpler and low-cost detection methods
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