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Production of highly sensitive monoclonal antibody and development of lateral flow assays for phallotoxin detection in urine

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Abstract

Phallotoxins, toxic cyclopeptides found in wild poisonous mushrooms, are predominant causes of fatal food poisoning. For the early and rapid diagnosis mushroom toxin poisoning, a highly sensitive and robust monoclonal antibody (mAb) against phallotoxins was produced for the first time. The half-maximum inhibition concentration (IC50) values of the mAb-based indirect competitive ELISAs for phallacidin (PCD) and phalloidin (PHD) detection were 0.31 ng mL−1 and 0.35 ng mL−1, respectively. In response to the demand for rapid screening of the type of poisoning and accurate determination of the severity of poisoning, colloidal gold nanoparticle (GNP) and time-resolved fluorescent nanosphere (TRFN) based lateral flow assays (LFA) were developed. The GNP-LFA has a visual cut-off value of 3.0 ng mL−1 for phallotoxins in human urine sample. The TRFN-LFA provides a quantitative readout signal with detection limit of 0.1 ng mL−1 in human urine sample. In this study, urine samples without pretreatment were used directly for the LFA strip tests, and both two LFAs were able to accomplish analysis within 10 min. The results demonstrated that LFAs based on the newly produced, highly sensitive, and robust mAb were able to be used for both rapid qualitative screening of the type of poisoning and accurate quantitative determination of the severity of poisoning after accidental ingestion by patients of toxic mushrooms.

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Funding

This work was supported by the Chinese Ministry of Science and Technology (MOST) for the National Key R&D Program of China (2018YFC1602600).

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Authors and Affiliations

  1. College of Veterinary Medicine, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety Beijing Laboratory for Food Quality and Safety, China Agricultural University, 100193, Beijing, People’s Republic of China

    Jianyu Zhu, Leina Dou, Jiafei Mi, Yuchen Bai, Minggang Liu, Jianzhong Shen, Wenbo Yu, Suxia Zhang, Xuezhi Yu & Zhanhui Wang

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  1. Jianyu Zhu
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  2. Leina Dou
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  3. Jiafei Mi
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  4. Yuchen Bai
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  7. Wenbo Yu
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  8. Suxia Zhang
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  10. Zhanhui Wang
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Correspondence to Suxia Zhang or Xuezhi Yu.

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Ethics approval

Urine samples were obtained directly from volunteers, having given their informed consent for collection. The studies have been approved by Ethics Committee of Shenzhen Center for Disease Control and Prevention (Research Ethical Review Number R2018021) and have been performed in accordance with the ethical standards.

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Zhu, J., Dou, L., Mi, J. et al. Production of highly sensitive monoclonal antibody and development of lateral flow assays for phallotoxin detection in urine. Anal Bioanal Chem 413, 4979–4987 (2021). https://doi.org/10.1007/s00216-021-03457-7

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  • DOI: https://doi.org/10.1007/s00216-021-03457-7

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