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

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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 105 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.

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Acknowledgments

This research was supported by the Capability Construction Program of Science and Technology Commission of Shanghai (13430502400), Science and Technology Innovation Plan of Shanghai: Yangtze River Delta Joint Research (15395810900), Postgraduate Education Innovation Program of Shanghai and the Special Fund for Agro-Scientific Research in the Public Interest (201003066).

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    Authors and Affiliations

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

      Haijuan Zeng, Jianwu Li, Xuzhao Zhai, Chunmei Song & Qing Liu

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

      Wenbo Guo

    3. Shanghai University of Medicine and Health Sciences, Shanghai, 200093, China

      Beibei Liang

    4. Chinese Academy of Inspection and Quarantine, Beijing, 100029, China

      Wenjun Zhao

    5. Institute of Biochemistry and Molecular Biology, ZBMZ, University of Freiburg, 79104, Freiburg, Germany

      Enguo Fan

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    1. Haijuan Zeng
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    Correspondence to Wenjun Zhao, Enguo Fan or Qing Liu.

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    All animal studies have been approved by China Ethics Committee and performed in accordance with the ethical standards.

    All of the authors agree to the submission of this paper. The corresponding authors declare on behalf of their co-authors that the work described was original research that has not been published previously, and not under consideration for publication elsewhere, in whole or in part.

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    Haijuan Zeng and Wenbo Guo contributed equally to this work.

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    Zeng, H., Guo, W., Liang, B. et al. Self-paired monoclonal antibody lateral flow immunoassay strip for rapid detection of Acidovorax avenae subsp. citrulli . Anal Bioanal Chem 408, 6071–6078 (2016). https://doi.org/10.1007/s00216-016-9715-5

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