水产品中环境雌激素生物检测技术的开发与应用
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- 英文论文题名：Development and application of a bioassay for detecting environmental estrogens in aquatic products using vitellogenin in Limanda yokohamae
- 论文作者：王军 ; 王骏 ; 张振忠 ; 汝少国
- 英文论文作者：WANG Jun ; WANG Jun ; ZHANG Zhenzhong ; RU Shaoguo ; Marine Life Science College ; Ocean University of China ; Shandong Entry-Exit Inspection and Quarantine Bureau
- 年：2016
- 作者机构：中国海洋大学海洋生命学院;山东省出入境检验检疫局;
- 论文关键词：环境雌激素 ; 黄盖鲽 ; 卵黄原蛋白 ; 夹心ELISA
- 英文论文关键词：Environmental estrogen ; Limanda yokohamae ; Vitellogenin ; Sandwich ELISA
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：X836
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：573k
- 原文格式：D
- 会议级别：全国
摘要
水体中的雌激素类污染物可以通过生物富集效应与食物链放大作用对人体健康产生潜在危害,已引起广泛关注。本研究利用卵黄原蛋白(Vitellogenin,Vtg)建立了水产品中环境雌激素的生物标志物检测技术,并以具有雌激素活性的全氟辛酸为目标污染物检验了方法的可靠性。结果显示,凝胶过滤与离子交换层析方法从17β-雌二醇诱导后的黄盖鲽(Limanda yokohamae)血浆中纯化的Vtg是一种分子量约为545 kDa的糖磷脂蛋白,在SDS-PAGE显示分子量约为162与108 kDa的主带;通过免疫大白兔制备了高特异性的Vtg多克隆抗体,以纯化的Vtg抗体包被酶标板,以HRP标记的Vtg抗体为二抗,建立了Vtg的夹心ELISA,其工作范围为3.9~500 ng/mL,检出限为1.5 ng/mL,组内与组间变异系数均小于10%。利用建立的夹心ELISA测定了全氟辛酸暴露后雄性黄盖鲽血浆中的Vtg含量,发现Vtg浓度随鱼肉中蓄积的全氟辛酸含量升高而增加,表明Vtg指标可以有效地指示全氟辛酸在鱼体内的蓄积情况。该研究初步证实鱼类VtgELISA可用作水产品中环境雌激素的生物检测技术。
Environmental estrogens have aroused much concern for their potential harm to human and wildlife. In the present stay, we developed and validated a method for detecting environmental estrogens based on vitellogenin(Vtg). Vtg purified from the plasma of 17β-estradiol induced Limanda yokohamae was characterized and used to prepare polyclonal antibody. Using the purified Vtg, anti-Vtg antibody, and HRP-labeled anti-Vtg antibody, a sandwich ELISA for quantifying Vtg was developed. This assay showed high sensitivity, specificity, and robustness.Furthermore, the reliability of the ELISA for detecting exogenous estrogens was confirmed by quantifying plasma Vtg in Limanda yokohamae exposed to Perfluorooctanoic acid(PFOA). We found that Vtg concentrations in plasma increased with the levels of PFOA accumulated in muscle tissues, indicating that Vtg ELISA could be used as a biological detection method for estrogenic contamination in aquatic products.
Environmental estrogens have aroused much concern for their potential harm to human and wildlife. In the present stay, we developed and validated a method for detecting environmental estrogens based on vitellogenin(Vtg). Vtg purified from the plasma of 17β-estradiol induced Limanda yokohamae was characterized and used to prepare polyclonal antibody. Using the purified Vtg, anti-Vtg antibody, and HRP-labeled anti-Vtg antibody, a sandwich ELISA for quantifying Vtg was developed. This assay showed high sensitivity, specificity, and robustness.Furthermore, the reliability of the ELISA for detecting exogenous estrogens was confirmed by quantifying plasma Vtg in Limanda yokohamae exposed to Perfluorooctanoic acid(PFOA). We found that Vtg concentrations in plasma increased with the levels of PFOA accumulated in muscle tissues, indicating that Vtg ELISA could be used as a biological detection method for estrogenic contamination in aquatic products.
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