High-Sensitivity Express Immunochromatographic Method for Detection of Plant Infection by Tobacco Mosaic Virus

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Yu. F. Drygin^1*, A. N. Blintsov², A. P. Osipov³, V. G. Grigorenko³, I. P. Andreeva⁴, A. I. Uskov⁵, Yu. A. Varitsev⁵, B. V. Anisimov⁵, V. K. Novikov^2#, and J. G. Atabekov^1,2

¹Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991 Moscow, Russia; fax: (495) 939-3181; E-mail: drygin@belozersky.msu.ru

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

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

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

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

^# Deceased.

^* To whom correspondence should be addressed.

Received February 5, 2009; Revision received March 11, 2009

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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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KEY WORDS: tobacco mosaic virus, immunochromatography analysis, colloidal gold, lateral flow immune analysis

DOI: 10.1134/S0006297909090065

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