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

Yu. F. Drygin1*, A. N. Blintsov2, A. P. Osipov3, V. G. Grigorenko3, I. P. Andreeva4, A. I. Uskov5, Yu. A. Varitsev5, B. V. Anisimov5, V. K. Novikov2#, and J. G. Atabekov1,2

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

DOI: 10.1134/S0006297909090065