Development of a fast ELISA for the specific detection of both leucomalachite green and malachite green

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• 作者：Yousheng Jiang ; Li Chen ; Kun Hu ; Wenjuan Yu…
• 关键词：monoclonal antibody ; Malachite Green ; ELISA ; fish
• 刊名：Journal of Ocean University of China
• 出版年：2015
• 出版时间：April 2015
• 年：2015
• 卷：14
• 期：2
• 页码：340-344
• 全文大小：316 KB
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• 刊物主题：Oceanography; Meteorology;
• 出版者：Springer Berlin Heidelberg
• ISSN：1993-5021

文摘

Malachite green (MG), a dye, is an antifungal agent that has been used to treat and prevent fish diseases. It is metabolized into reduced leucomalachite green forms (LMG) that may reside in fish muscles for a long period, thus being harmful to human health. The aim of this study was to develop a competitive and direct enzyme-linked immunosorbent assay (ELISA) to detect MG and LMG specifically. The monoclonal antibody (mAb) to LMG was generated using a hybridoma technique. The obtained mAb showed good cross-reactivity (CR) to malachite green (MG), but not to crystal violet (CV) and Brilliant Green (BG). The mAb was used to develop a fast detecting ELISA of MG and LMG in fish. By introducing the conjugation LMG-HRP, the detection capability was 0.37 ng mL? for MG and LMG. The mean recovery from spiked grass carp tissues ranged from 76.2% to 82.9% and the coefficients of variation varied between 1.8% and 7.5%. The stable and efficient monoclonal cell line obtained is a sustainable source of sensitive and specific antibody to MG and LMG.