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Development of a lateral flow immunoassay for rapid diagnosis of potato blackleg caused by Dickeya species

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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 × 105 CFU/ml for leaves and 4 × 105 CFU/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.

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

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Acknowledgements

The study was financially supported by the Russian Science Foundation (grant no. 16-16-04108).

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Authors and Affiliations

  1. A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospekt 33, Moscow, 119071, Russia

    Irina V. Safenkova, Nadezhda A. Byzova, Anatoly V. Zherdev & Boris B. Dzantiev

  2. A.G. Lorch All-Russian Potato Research Institute, Kraskovo-1, Moscow Region, 140051, Russia

    Ilya A. Zaitsev & Yuri A. Varitsev

  3. All-Russian Plant Quarantine Center, Bykovo-2, Moscow Region, 140150, Russia

    Natalia V. Drenova

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  1. Irina V. Safenkova
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  3. Yuri A. Varitsev
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  4. Nadezhda A. Byzova
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  5. Natalia V. Drenova
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  6. Anatoly V. Zherdev
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  7. Boris B. Dzantiev
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Correspondence to Boris B. Dzantiev.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Safenkova, I.V., Zaitsev, I.A., Varitsev, Y.A. et al. Development of a lateral flow immunoassay for rapid diagnosis of potato blackleg caused by Dickeya species. Anal Bioanal Chem 409, 1915–1927 (2017). https://doi.org/10.1007/s00216-016-0140-6

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