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烟草花叶病毒感染植物的高灵敏度 express 免疫层析检测方法。

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

机构信息

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

出版信息

Biochemistry (Mosc). 2009 Sep;74(9):986-93. doi: 10.1134/s0006297909090065.

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.

摘要

一种用于植物病毒感染分子诊断的高灵敏度免疫层析法,以烟草花叶病毒(TMV)为例进行了阐述。分析时间不超过 5 分钟,在未澄清的叶片提取物中检测 TMV 的下限(2-4ng/ml)与病毒的酶联免疫吸附测定的灵敏度相当。单次测量需要 0.1-0.2ml 测试溶液(从 10-20mg 叶片材料中提取)。使用银的信号增强,将 TMV 在叶片组织提取物中的测定灵敏度提高了一个数量级以上,达到 0.1ng/ml。在这种情况下,分析时间不超过 25 分钟。该方法的简单性使其在大量分析样本的快速诊断中尤为方便。阐述了一种用于实验室和田间条件下植物病毒感染连续分析的诊断试剂盒的原型。

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