Fish Endocrinology Laboratory, Department of Zoology, Banaras Hindu University, Varanasi, 221 005, India,
Fish Physiol Biochem. 2005 Apr;31(2-3):289-93. doi: 10.1007/s10695-006-0038-x.
This study describes the development and validation of competitive antigen capture enzyme-linked immunosorbent assay (ELISA) for growth hormone (GH) of the catfish, Clarias batrachus. Isolated GH was characterized first through bioassay, amino acid sequencing, immunoblotting and immunocytochemistry, and then used to raise antibody to develop ELISA. The lowest detection limit of the assay system was 0.17 ng ml(-1), and the standard curve had an ED(50) value of 0.35 ng ml(-1). Repeated determination of GH in a plasma pool exhibited intra- and interassay co-efficient of variation of 8.1% and 7.09% (n=5), respectively. Dose-response inhibition curves resulting from dilutions of plasma and pituitary homogenate of C. batrachus were parallel to the standard curve, while such parallelism in the case of Cyprinus carpio and H. fossilis was absent revealing no cross reaction of them in the present ELISA. In vivo effects of sGnRH, testosterone, 5-HT and morphine on plasma GH were also examined to validate the ELISA physiologically. sGnRH, testosterone and morphine increased, while 5-HT decreased GH level in a dose-dependent manner.
本研究描述了鲶鱼（Clarias batrachus）生长激素（GH）的竞争性抗原捕获酶联免疫吸附测定（ELISA）的开发和验证。首先通过生物测定、氨基酸测序、免疫印迹和免疫细胞化学对分离出的 GH 进行了表征，然后用其来产生抗体以开发 ELISA。该测定系统的最低检测限为 0.17ng/ml，标准曲线的 ED（50）值为 0.35ng/ml。在对血浆池进行重复测定时，日内和日间变异系数分别为 8.1%和 7.09%（n=5）。鲶鱼血浆和脑垂体匀浆稀释产生的剂量-反应抑制曲线与标准曲线平行，而在鲤鱼和纹缟𬶐的情况下则不存在平行性，表明在本 ELISA 中它们之间没有交叉反应。还通过体内实验验证了 sGnRH、睾酮、5-HT 和吗啡对血浆 GH 的生理影响。sGnRH、睾酮和吗啡呈剂量依赖性增加 GH 水平，而 5-HT 则降低 GH 水平。
