基于碱性磷酸酶.pdfVIP

基于碱性磷酸酶-金刚烷体系的化学发光酶免疫分析法对磺胺对甲氧 嘧啶的测定 梁艳利a a b* b a ，闫娜利 ，吴拥军 ，玉崧成 ，屈凌波 (郑州大学，a.化学系，b.公共卫生学院，郑州 450001) 摘要：目的 用碱性磷酸酶(ALP)-金刚烷(AMPPD)体系，建立化学发光酶免疫分析定量检测磺胺对甲氧嘧啶(SMD)的方 法。方法 利用重氮化偶联法标记 ALP ，在优化的实验条件下抗体包被浓度为 5.0 μg·mL1 ，酶标记物工作液稀释度为 1 ∶10，每孔加待测样品或标准品50 μL 和酶标记物50 μL ，在37 ℃下反应l h ，洗涤后每孔加发光底物100 μL ，避光反 应25 min 后在发光检测仪下检测发光计数。结果 该方法检测限为28.0 pg·mL1 ，批内变异系数为13.5%~14.6%，批间 作者简介：梁艳利，女，硕士生 Tel:E-mail: liangyanli3122005@163.com *通信作者：吴拥军，男，博士，教授 Tel: (0371E-mail: wuyongjun@ ·550 · Chin JMAP, 2011 June, Vol.28 No.6 中国现代应用药学2011 年6 月第28 卷第6 期 变异系数为16.1%~17.5%。标准曲线回归方程为Y=0.021 5X +5.172 6(r=0.996 1) ，线性范围为50~10 000 pg·mL1。结论 首 次运用ALP-AMPPD 发光体系的化学发光酶免疫分析法实现了对SMD 残留的快速检测。 关键词：碱性磷酸酶；金刚烷；化学发光酶免疫分析；磺胺对甲氧嘧啶 中图分类号：R927.2 文献标志码：B 文章编号：1007-7693(2011)06-0550-04 Determination of Sulfameter by Chemiluminescence Enzyme Immunoassay Based on Alkaline Phosphatase-Adamantane a a b* b a LIANG Yanli , YAN Nali , WU Yongjun , YU Songcheng , QU Lingbo (Zhengzhou University, a.Department of Chemistry, b.College of Public Health, Zhengzhou 450001, China) ABSTRACT: OBJECTIVE To establish a chemiluminescence enzyme immunoassay detection method for the quantitative assay of sulfameter, using alkaline phosphatase (ALP)-adamantane (AMPPD) system. METHODS The diazotization-coupling method was used to combine horseradish peroxidase with sulfameter. The optimal conditions were as follows. The concentration of antibody-coated solid-phase was 5.0 μg·mL1. The ALP-sulfameter was diluted to l ∶10. Every well was added by standards or samples (50 μL) and ALP-sulfameter (50 μL), then plates were placed at 37 ℃for 1 h. After washed, each well was added by 100 μL AMPPD. The result was obtained by luminometer after adding substrate for 25 min.