Jensen Kathleen M, Ankley Gerald T
National Health and Environmental Effects Research Laboratory, Office of Research and Development, Mid-Continent Ecology Division, US Environmental Protection Agency, 6201 Congdon Boulevard, Duluth, MN 55804, USA.
Ecotoxicol Environ Saf. 2006 Jun;64(2):101-5. doi: 10.1016/j.ecoenv.2006.02.011. Epub 2006 Apr 17.
Vitellogenin (vtg) concentrations in oviparous animals such as fish represent an integrated indicator of the status of the reproductive endocrine system. As such, vtg is a common measurement endpoint in tests designed to detect certain classes of endocrine-disrupting chemicals (EDCs). The most common approach to measuring vtg is via enzyme-linked immunosorbent assays (ELISAs). However, because labs testing EDCs in fish often use slightly different ELISAs (e.g., in terms of antibodies, binding antigens, standards), results among studies are not always comparable. One approach to obviating this would be for researchers to use standardized ELISA kits from a common source(s). The fathead minnow (Pimephales promelas) is a small fish model commonly used for EDC testing. The purpose of this study was to evaluate a recently developed commercial ELISA kit for measuring vtg in the fathead minnow. The commercial ELISA, based on a monoclonal antibody to fathead minnow vtg, was compared to an ELISA that utilizes a fathead minnow polyclonal antibody, which has been used extensively in our lab and others for several years. Plasma samples for this comparison came from three studies in which fathead minnows had been exposed to different model EDCs, including an androgen (17beta-trenbolone), an anti-androgen (flutamide), and two CYP19 (aromatase) inhibitors (prochloraz, fadrozole). Results obtained using the two different ELISA methods were consistently similar.
在鱼类等卵生动物中,卵黄蛋白原(vtg)浓度是生殖内分泌系统状态的综合指标。因此,在旨在检测某些类别的内分泌干扰化学物质(EDC)的测试中,vtg是一个常见的测量终点。测量vtg最常用的方法是酶联免疫吸附测定(ELISA)。然而,由于检测鱼类中EDC的实验室通常使用略有不同的ELISA(例如,在抗体、结合抗原、标准品方面),不同研究的结果并不总是具有可比性。避免这种情况的一种方法是研究人员使用来自同一来源的标准化ELISA试剂盒。黑头呆鱼(Pimephales promelas)是一种常用于EDC测试的小型鱼类模型。本研究的目的是评估一种最近开发的用于测量黑头呆鱼中vtg的商业ELISA试剂盒。将基于针对黑头呆鱼vtg的单克隆抗体的商业ELISA与一种使用黑头呆鱼多克隆抗体的ELISA进行比较,该多克隆抗体已在我们实验室和其他实验室广泛使用数年。用于此次比较的血浆样本来自三项研究,在这些研究中,黑头呆鱼暴露于不同的模型EDC中,包括一种雄激素(17β-群勃龙)、一种抗雄激素(氟他胺)和两种CYP19(芳香化酶)抑制剂(咪鲜胺、法倔唑)。使用两种不同ELISA方法获得的结果始终相似。