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用于快速检测西瓜嗜酸菌的自配对单克隆抗体侧流免疫分析试纸条

Self-paired monoclonal antibody lateral flow immunoassay strip for rapid detection of Acidovorax avenae subsp. citrulli.

作者信息

Zeng Haijuan, Guo Wenbo, Liang Beibei, Li Jianwu, Zhai Xuzhao, Song Chunmei, Zhao Wenjun, Fan Enguo, Liu Qing

机构信息

School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai, 200093, China.

Institute of Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, Shanghai, 201403, China.

出版信息

Anal Bioanal Chem. 2016 Sep;408(22):6071-8. doi: 10.1007/s00216-016-9715-5. Epub 2016 Jul 1.

Abstract

We screened a highly specific monoclonal antibody (McAb), named 6D, against Acidovorax avenae subsp. citrulli (Aac). Single McAb 6D was used as both nanogold-labeled antibody and test antibody to develop a single self-paired colloidal gold immunochromatographic test strip (Sa-GICS). The detection limit achieved using the Sa-GICS approach was 10(5) CFU/mL, with a result that can be observed by the naked eye within 10 min. Moreover, Sa-GICS can detect eight strains of Aac and display no cross-reactions with other pathogenic plant microorganisms. Artificial contamination experiments demonstrated that Sa-GICS would not be affected by impurities in the leaves or stems of the plants and were consistent with the PCR results. This is the first report on the development of a colloidal gold immunoassay strip with self-paired single McAb for the rapid detection of Aac. Graphical Abstract Schematic representation of the test strip.

摘要

我们筛选了一种针对西瓜嗜酸菌(Aac)的高度特异性单克隆抗体(McAb),名为6D。单一的单克隆抗体6D既用作纳米金标记抗体,又用作检测抗体,以开发一种单自配对胶体金免疫层析试纸条(Sa-GICS)。使用Sa-GICS方法达到的检测限为10⁵ CFU/mL,结果可在10分钟内用肉眼观察到。此外,Sa-GICS可以检测八株Aac菌株,并且与其他致病植物微生物无交叉反应。人工污染实验表明,Sa-GICS不会受到植物叶片或茎干中杂质的影响,并且与PCR结果一致。这是关于开发用于快速检测Aac的具有自配对单克隆抗体的胶体金免疫分析试纸条的首次报道。图形摘要试纸条的示意图。

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