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Lateral flow immunoassay for rapid detection of potato ring rot caused by Clavibacter michiganensis subsp. sepedonicus

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Abstract

A lateral flow immunoassay for the rapid detection of Clavibacter michiganensis subsp. sepedonicus bacteria causing potato ring rot was developed. Multimembrane composites (test strips) containing polyclonal antibodies against the bacteria and gold nanoparticle-antibody conjugates were used for the analysis. The test strips are suitable for the analysis of potato tuber and leaf extracts within 10 min; the detection limit of bacteria is 4 × 105 cells/mL. No cross-reactivity with strains of Clavibacter michiganensis subsp. michiganensis, Pectobacterium carotovorum subsp. carotovorum and saprophytes of healthy potato plants was detected. The results of analysis of 26 potato samples by the developed tests were compared with those obtained by the PCR method and using the commercial enzyme immunoassay kits. The results of lateral flow immunoassay were confirmed in 96.2% of cases, which supports the high correlation with other analytical approaches. The developed immunoassay may be considered as a promising means of phytosanitary control.

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Abbreviations

Cmm:

Clavibacter michiganensis subsp. michiganensis

Cms:

Clavibacter michiganensis subsp. sepedonicus

ELISA:

enzyme-linked immunosorbent assay

GNP:

gold nanoparticles

IgG:

immunoglobulin G

LFIA:

lateral flow immunoassay

PBS:

phosphate buffered saline (50 mM K-phosphate buffer, pH 7.4, 0.14 M NaCl)

PBST:

PBS with 0.05% Triton X-100

PCR:

polymerase chain reaction

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Authors and Affiliations

  1. A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, 119071, Russia

    I. V. Safenkova, G. K. Pankratova, A. V. Zherdev & B. B. Dzantiev

  2. All-Russian Potato Research Institute, Kraskovo-1, Moscow region, 140051, Russia

    I. A. Zaitsev & Yu. A. Varitsev

Authors
  1. I. V. Safenkova
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  2. I. A. Zaitsev
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  3. G. K. Pankratova
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  4. Yu. A. Varitsev
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  5. A. V. Zherdev
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  6. B. B. Dzantiev
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Corresponding author

Correspondence to B. B. Dzantiev.

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Safenkova, I.V., Zaitsev, I.A., Pankratova, G.K. et al. Lateral flow immunoassay for rapid detection of potato ring rot caused by Clavibacter michiganensis subsp. sepedonicus . Appl Biochem Microbiol 50, 675–682 (2014). https://doi.org/10.1134/S0003683814120011

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  • DOI: https://doi.org/10.1134/S0003683814120011

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