Development of ELISAs for the detection of vitellogenin in three marine fish from coastal areas of China
Development of ELISAs for the detection of vitellogenin in three marine fish from coastal areas of China
复制标题
建立检测中国沿海三种海水鱼类卵黄蛋白原的 ELISA 方法
DOI：
10.1016/j.marpolbul.2018.06.001
复制
发表时间：
2018-08-01
影响因子：
5.8
通讯作者：
Ru, Shaoguo
中科院分区：
文献类型：
--
作者：
Li, Yuejiao;Wang, Jun;Ru, Shaoguo
雌激素污染对海洋生物的危害已引起人们的广泛关注。本研究旨在建立基于生物标志物的中国沿海硬骨鱼类卵黄蛋白原（Vtg）环境雌激素检测方法。首先，对花鲽、许氏平鲉和大泷石斑鱼的Vtgs进行了纯化、鉴定和抗体制备。然后，使用纯化的Vtg及其抗体开发了每个物种的Vtg ELISA。大理石鲽Vtg ELISA的工作范围为3.9 ~ 500 ng/mL，检出限为2.1 ng/mL，黑石斑鱼Vtg ELISA与大理石鲽Vtg具有较强的交叉反应性。此外，Vtg的诱导暴露于十五碳氟辛酸（PFOA）的男性大理石花纹比目鱼测定发达的ELISA。随着海水和肌肉中PFOA浓度的增加，血浆Vtg浓度显著增加。因此，这些物种的Vtg ELISA可能是有用的工具，监测海洋环境雌激素。
Estrogenic pollution has aroused great concern for its adverse effects on marine organisms. This study aimed to establish biomarker-based methods for detecting environmental estrogens using vitellogenin (Vtg) of teleost fishes inhabiting coastal areas of China. Firstly, Vtgs in marbled flounder (Pseudopleuronectes yokohamae), black rockfish (Sebastes schlegelii) and fat greenling (Hexagrammos otakii) were purified, characterized and used to prepare antibodies. Then, Vtg ELISA for each species was developed using purified Vtg and its antibody. Marbled flounder Vtg ELISA had a working range of 3.9-500 ng/mL and a detection limit of 2.1 ng/mL, and black rockfish Vtg ELISA had strong cross-reactivity with marbled flounder Vtg. Furthermore, Vtg induction in male marbled flounder exposed to pentadecafluorooctanoic acid (PFOA) was measured by developed ELISA. Plasma Vtg concentrations were significantly increased with PFOA concentrations in seawater and fish muscle. Therefore, Vtg ELISAs for these species might be useful tools for monitoring marine environmental estrogens.