title: Development of a lateral flow immunoassay for rapid diagnosis of potato blackleg caused by Dickeya species.
abstract: Early detection of potato infections is essential for effective disease management. The aim of this study was to develop a lateral flow immunoassay (LFIA) for rapid detection of a serious potato disease, potato blackleg, caused by Dickeya dianthicola and Dickeya solani. Polyclonal antibodies specific to different strains of Dickeya were obtained from rabbits after immunization with bacterial cells of D. dianthicola and D. solani. Enzyme-linked immunosorbent assay testing with use of a wide range of bacterial species showed that the polyclonal antibodies detect closely related strains of D. dianthicola and D. solani. Cross-reactivity with widespread pathogenic bacteria (nine species) and saprophytes of healthy potato plants was not detected. The LFIA based on the obtained antibodies and gold nanoparticles with average diameter of 20 nm was developed. Under optimized conditions, the LFIA method enabled the analysis of potato extracts within 10 min, with a visual limit of detection of 1 × 105CFU/ml for leaves and 4 × 105CFU/ml for tubers. The assay was tested on potato stem and tuber extracts, and the results of the LFIA were confirmed in 92.1% of samples using the real-time polymerase chain reaction. The findings confirmed that the developed LFIA could be used for monitoring blackleg infection without the need for special equipment or skills. Graphical Abstract The developed lateral flow immunoassay is an efficient tool for rapid detection of a serious potato disease, potato blackleg, caused by Dickeya dianthicola and Dickeya solani.
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospekt 33, Moscow, 119071, Russia.
authors.forename: Irina V
authors.initials: IV
authors.lastname: Safenkova
authors.affiliation: A.G. Lorch All-Russian Potato Research Institute, Kraskovo-1, Moscow Region, 140051, Russia.
authors.forename: Ilya A
authors.initials: IA
authors.lastname: Zaitsev
authors.affiliation: A.G. Lorch All-Russian Potato Research Institute, Kraskovo-1, Moscow Region, 140051, Russia.
authors.forename: Yuri A
authors.initials: YA
authors.lastname: Varitsev
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospekt 33, Moscow, 119071, Russia.
authors.forename: Nadezhda A
authors.initials: NA
authors.lastname: Byzova
authors.affiliation: All-Russian Plant Quarantine Center, Bykovo-2, Moscow Region, 140150, Russia.
authors.forename: Natalia V
authors.initials: NV
authors.lastname: Drenova
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospekt 33, Moscow, 119071, Russia.
authors.forename: Anatoly V
authors.initials: AV
authors.lastname: Zherdev
authors.affiliation: A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospekt 33, Moscow, 119071, Russia. dzantiev@inbi.ras.ru.
authors.forename: Boris B
authors.initials: BB
authors.lastname: Dzantiev
mesh_terms: D000906:Antibodies / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D020563:Gammaproteobacteria / Q000276:immunology / Q000472:pathogenicity*; D007118:Immunoassay / Q000379:methods*; D046529:Microscopy, Electron, Transmission; D010935:Plant Diseases / Q000382:microbiology*; D011198:Solanum tuberosum / Q000382:microbiology*
chemical_list: D000906:Antibodies
keywords: Dickeya spp.; Gold nanoparticles; Lateral flow immunoassay; Phytopathogens