| 竞争性ELISA法检测人尿金属硫蛋白方法的改进及应用 |
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| 引用本文： | 贾连群,葛正龙,金泰,包意芳,欧刚卫.竞争性ELISA法检测人尿金属硫蛋白方法的改进及应用[J].劳动医学,2010,27(3):131-134. |
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| 作者姓名： | 贾连群 葛正龙 金泰 包意芳 欧刚卫 |
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| 作者单位： | 辽宁中医药大学基础医学院;遵义医学院生物化学与分子生物学教研室;复旦大学公共卫生学院劳动卫生教研室;中国疾病预防控制中心寄生虫病研究所; |
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| 基金项目： | 遵义医学院院基金资助(编号:F070) |
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| 摘 要： | 目的]改进检测人尿金属硫蛋白（UMT）的竞争性酶联免疫吸附法（ELISA），探讨应用该法测定人尿样标本UMT含量的价值。方法]分离纯化人肝脏金属硫蛋白（HMT），制备抗金属硫蛋白单克隆抗体（McAb），利用待测样品中MT与包被MT竞争结合McAb的原理，建立竞争性ELISA，并检测健康在校大学生298份尿样中的UMT含量。结果]竞争性ELISA对人UMT最低检出限为9ng／mL，组内和组间的变异系数分别为1．78％一6．72％和5．45％～13：26％，样品的回收率为99．69％～107．25％。尿样检测结果显示运动后UMT含量为（19．73±8．65）ng／μmolCr较运动前的（10．58±5．62）ng／μmolCr明显增加（P〈0．05）；吸烟群体UMT含量为（86．58±30．96）ng／μmolCr，显著高于不吸烟人群（14．73±6．57）ng／μmolCr]（P〈0．05）。结论]本法对于检测人UMT具有较好的精密度和准确度，可用于人UMT检测；运动后UMT水平显著升高，吸烟者UMT明显高于正常对照。
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| 关 键 词： | 尿金属硫蛋白 单克隆抗体 酶联免疫吸附测定 |
Improvement and Application of a Competitive ELISA for Detection of Metallothionein in Human Urine |
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| JIA Lian-qun,GE Zheng-long,JIN Tai-yi,BAO Yi-fang,OU Gang-wei.Improvement and Application of a Competitive ELISA for Detection of Metallothionein in Human Urine[J].Journal of Labour Medicine,2010,27(3):131-134. |
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| Authors: | JIA Lian-qun GE Zheng-long JIN Tai-yi BAO Yi-fang OU Gang-wei |
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| Institution: | 1.Department of Biochemistry and Molecular Biology/a>;Liaoning University of Traditional Chinese Medicine/a>;Shenyang/a>;Liaoning 110032/a>;China/a>;2.Department of Biochemistry and Molecular Biology/a>;Zunyi Medical College/a>;Zunyi/a>;Guizhou 563003/a>;3.Department of Occupational Health/a>;School of Public Health/a>;Fudan University/a>;Shanghai 200032/a>;4.National Institute of Parasitic Diseases/a>;Chinese Center for Disease Control and Prevention 200025/a>;China |
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| Abstract: | Objective]To improve and apply the competitive ELISA method for measuring human urinary metallothionein(UMT). Methods] Human metallothionein(MT)was isolated from an adult liver by SephadexG-75 and Sephadex G-50 chromatographic column,and then used to immunize Balb/c mice for preparing monoclonal antibody(McAb).The competitive ELISA was established with the McAb as the primary antibody.UMT was determined in 298 urine samples from university students.Results] The detection limit of the ELISA was 9 ng/mL.Th... |
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| Keywords: | metallothionein monoclonal antibody competitive ELISA |
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