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首页 > 过刊浏览>2023年第23卷第11期 >353-360. DOI:10.16429/j.1009-7848.2023.11.033
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水产品中氯霉素酶联免疫吸附分析法的构建与应用
作者:
作者单位:

(1.中国计量大学生命科学学院 杭州 310018;2.杭州铭治医学检验实验室有限公司 杭州 310020)

基金项目:

浙江省重点研发计划项目(2022C02049，2021C02061)


Construction and Application of Enzyme-linked Immunosorbent Assay for Chloramphenicol in Aquatic Products
Author:
Affiliation:

(1.School of Life Sciences， China Jiliang University， Hangzhou 310018;2.Hangzhou Mingzhi Medical Laboratory Co. Ltd.， Hangzhou 310020)

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    摘要:

    以氯霉素(CAP)为研究对象，通过系统优化反应条件，构建CAP的间接竞争酶联免疫分析法，并应用于水产品中CAP残留量的检测。氯霉素间接竞争酶联免疫分析方法的最佳工作条件为:0.05 μg/孔包被量(CAP-OVA)，4 ℃包被14 h，0.5%脱脂乳粉封闭液，CAP抗体稀释倍数为1∶16 000。结果表明:构建的间接竞争酶联免疫分析方法的灵敏度(IC50)为4.88 μg/mL，检测限(IC15)为0.29 μg/mL，方法特异性良好，对氯霉素结构类似物和同类抗生素的交叉率低于0.1%。对大黄鱼、缢蛏、牛蛙进行5.0，10.0 μg/kg和20.0 μg/kg加标回收试验，回收率在83.90%~100.73%范围。本方法的日内和日间变异系数范围为2.28%~6.96%，该检测结果与LC-MS/MS分析结果具有良好的一致。本研究构建的CAP酶联免疫吸附分析法操作简单，灵敏度高，可应用于多种水产品中氯霉素残留量的快速定量检测，同时也为其它小分子危害物的酶联免疫分析方法的构建提供技术借鉴。

    Abstract:

    In this work， chloramphenicol (CAP) as the research object， by optimizing the reaction conditions， an indirect competitive enzyme-linked immunosorbent assay(ic-ELISA) for CAP was constructed and applied to the detection of CAP residues in aquatic products. The optimal working conditions of the constructed ic-ELISA were 0.05 μg/well coating amount of CAP-OVA， coating conditions at 4 ℃ for 14 h， 0.5% nonfat milk powder as blocking solution， and the dilution ratio of CAP antibody was 1∶16 000. The results showed that the sensitivity(IC50) of the constructed ic-ELISA method was 4.88 μg/mL， and the detection limit (IC15) was 0.29 μg/mL. The cross-reaction rate of the chloramphenicol structural analogs and similar antibiotics were less than 0.1%. The spiked recovery experiments of 5.0， 10.0， 20.0 μg/kg in Larimichthys crocea， razor clam and bullfrog were carried out， and the recovery rates were within 83.90%-100.73%， and the intraday and interday coefficients of variation (CV) for this method ranged from 2.28% to 6.96%. The detection results were in good agreement with that of LC-MS/MS. The ic-ELISA of CAP in this study was simple to operate and has high sensitivity. It could be applied to the rapid quantitative detection of CAP residues in various aquatic products， and provide technical reference for ic-ELISA of other small molecule hazards. And it is also an indirect competitive enzyme-linked immunosorbent assay for the construction of the method provides technical reference.

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张彪,蔡丹凤,姚天择,郎一涵,傅炎勇,申屠旭萍,俞晓平.水产品中氯霉素酶联免疫吸附分析法的构建与应用[J].中国食品学报,2023,23(11):353-360

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  • 收稿日期:2022-11-19
  • 在线发布日期: 2023-12-14
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