AN Belozersky Institute of Physico-chemical Biology, Moscow State University, Moscow, Russia.
Appl Microbiol Biotechnol. 2012 Jan;93(1):179-89. doi: 10.1007/s00253-011-3522-x. Epub 2011 Aug 26.
A test system is described and expanded upon for mass field immunochromatography assay on porous membrane carriers for rapid diagnostics of potato virus X (PVX) in potato leaf tissue and sprout extracts using colloidal gold nanoparticles as a marker. Sensitivity of the assay developed for PVX identification is found to be comparable to the sensitivity of solid-phase sandwich-ELISA. Complete assay time does not exceed 15 min, and the lower limit of the PVX detection in non-clarified leaf extract is 2 ng/ml. A single measurement requires 0.1-0.2 ml (3-5 drops) of tested solution only (extracted from 10-20 mg of potato leaf tissue or sprouts). The simplicity and reliability of the method makes it especially efficient in direct rapid monitoring of many infected potato specimens in the field, as verified by field trials of 360 clones of 28 domestic and foreign cultivars of potato. A diagnostic kit for routine analyses of potato viral infections both in the laboratory and in the field is described and expanded upon.
描述并扩展了一种测试系统,用于在多孔膜载体上进行大规模场免疫层析分析,以使用胶体金纳米颗粒作为标记物快速诊断马铃薯病毒 X(PVX)在马铃薯叶片组织和芽提取物中的存在。开发用于 PVX 鉴定的检测方法的灵敏度被发现与固相夹心 ELISA 的灵敏度相当。完整的检测时间不超过 15 分钟,在未澄清的叶片提取物中 PVX 的检测下限为 2ng/ml。单次测量仅需要 0.1-0.2 毫升(3-5 滴)测试溶液(从 10-20 毫克马铃薯叶片组织或芽中提取)。该方法的简单性和可靠性使其在田间直接快速监测许多受感染的马铃薯标本方面特别有效,这已通过对 28 个国内外马铃薯品种的 360 个克隆进行的田间试验得到验证。描述并扩展了一种用于实验室和田间常规分析马铃薯病毒感染的诊断试剂盒。