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High-sensitivity express immunochromatographic method for detection of plant infection by tobacco mosaic virus

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Abstract

A highly sensitive express immunochromatography method for molecular diagnosis of plant virus infections was elaborated on the example of a model object — tobacco mosaic virus (TMV). The analysis time does not exceed 5 min, and the lower limit of TMV detection in non-clarified leaf extract (2–4 ng/ml) is comparable with the sensitivity of the enzyme-linked immunosorbent assay of the virus. A single measurement requires 0.1–0.2 ml tested solution (extract from 10–20 mg of leaf material). The sensitivity of TMV determination in the leaf tissue extract was increased by more than one order of magnitude using signal enhancement by silver and is 0.1 ng/ml. In this case, analysis time did not exceed 25 min. The simplicity of this method makes it especially convenient in express diagnosis of numerous analyzed specimens. The prototype of a diagnostic kit for serial analyses of plant viral infections both in laboratory and field conditions was elaborated.

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Abbreviations

ABTS:

2,2′-azinobis(3-ethylbenzthiazolinesulfonic acid)

ELISA:

enzyme-linked immunosorbent assays

HRP:

horseradish peroxidase

IChA:

immunochromatography analysis

IgG-Au:

antibody conjugate with colloidal aurum

IgG-HRP:

antibody conjugate with horseradish peroxidase

TMV:

tobacco mosaic virus

TPB:

0.01 M K-phosphate buffer, pH 7.4, 0.1 M NaCl, 0.1% Triton X-100

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Authors and Affiliations

  1. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    Yu. F. Drygin & J. G. Atabekov

  2. Faculty of Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    A. N. Blintsov, V. K. Novikov & J. G. Atabekov

  3. Faculty of Chemistry, Lomonosov Moscow State University, 119991, Moscow, Russia

    A. P. Osipov & V. G. Grigorenko

  4. ZAO NVO Immunotech, Lomonosov Moscow State University, 119991, Moscow, Russia

    I. P. Andreeva

  5. All-Russian Potato Research Institute, ul. Lorkha 23, 140051, Kraskovo-1, Moscow Region, Russia

    A. I. Uskov, Yu. A. Varitsev & B. V. Anisimov

Authors
  1. Yu. F. Drygin
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  2. A. N. Blintsov
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  3. A. P. Osipov
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  4. V. G. Grigorenko
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  5. I. P. Andreeva
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  6. A. I. Uskov
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  7. Yu. A. Varitsev
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  8. B. V. Anisimov
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  9. V. K. Novikov
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  10. J. G. Atabekov
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Corresponding author

Correspondence to Yu. F. Drygin.

Additional information

Original Russian Text © Yu. F. Drygin, A. N. Blintsov, A. P. Osipov, V. G. Grigorenko, I. P. Andreeva, A. I. Uskov, Yu. A. Varitsev, B. V. Anisimov, V. K. Novikov, J. G. Atabekov, 2009, published in Biokhimiya, 2009, Vol. 74, No. 9, pp. 1212–1220.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM09-038, June 28, 2009.

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Drygin, Y.F., Blintsov, A.N., Osipov, A.P. et al. High-sensitivity express immunochromatographic method for detection of plant infection by tobacco mosaic virus. Biochemistry Moscow 74, 986–993 (2009). https://doi.org/10.1134/S0006297909090065

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  • DOI: https://doi.org/10.1134/S0006297909090065

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