基于一对特异性单克隆抗体的高灵敏度微孔板化学发光酶免疫分析法定量检测葡萄球菌肠毒素 B 及其在各种基质中的应用。
Highly sensitive microplate chemiluminescence enzyme immunoassay for the determination of staphylococcal enterotoxin B based on a pair of specific monoclonal antibodies and its application to various matrices.
机构信息
Department of Immunology, The Fourth Military Medical University, No. 17 Changle West Road, Xi'an 710032, Shaanxi Province, People's Republic of China.
出版信息
Anal Chem. 2010 Sep 15;82(18):7758-65. doi: 10.1021/ac101666y.
A highly specific and sensitive microplate chemiluminescent enzyme immunoassay (CLEIA) was established and validated for the detection of staphylococcal enterotoxin B (SEB). A pair of monoclonal antibodies (mAbs) that recognizes different epitopes of SEB was selected from 20 SEB-specific mAbs, and the experimental conditions were examined and optimized for the development of the CLEIA. This method exhibited high performance with a dynamic range of 0.01-5 ng/mL, and the measured limit of detection (LOD) was 0.01 ng/mL. Intra- and interassay coefficient variations were all lower than 13% at three concentrations (0.2, 0.4, and 2 ng/mL). For specificity studies, when this method was applied to test staphylococcal enterotoxins A, C1, and D, no cross-reactivity was observed. It has been successfully applied to the analysis of SEB in a variety of environmental, biological and humoral matrices such as sewage, tap water, river water, roast beef, peanut butter, cured ham, 10% nonfat dry milk, milk, orange juice, and human urine and serum. The aim of this article is to show that the highly sensitive, specific, and simple microplate CLEIA, based on a pair of highly specific monoclonal antibodies, has potential applications for quantifying SEB in public health and military reconnaissance.
建立并验证了一种用于检测葡萄球菌肠毒素 B（SEB）的高特异性和高灵敏度微孔板化学发光酶免疫分析（CLEIA）方法。从 20 株 SEB 特异性单克隆抗体（mAb）中筛选出识别 SEB 不同表位的一对 mAb，并对其进行了实验条件的检测和优化，以开发 CLEIA 方法。该方法具有 0.01-5ng/mL 的宽动态范围，测量的检测限（LOD）为 0.01ng/mL。在三个浓度（0.2、0.4 和 2ng/mL）下，批内和批间变异系数均低于 13%。特异性研究表明，当该方法用于检测葡萄球菌肠毒素 A、C1 和 D 时，没有观察到交叉反应。该方法已成功应用于污水、自来水、河水、烤牛肉、花生酱、熏火腿、10%脱脂奶粉、牛奶、橙汁和人尿和血清等多种环境、生物和体液基质中 SEB 的分析。本文旨在表明，基于一对高度特异性的单克隆抗体，这种高灵敏度、高特异性和简单的微孔板 CLEIA 方法在公共卫生和军事侦察中定量检测 SEB 具有潜在的应用价值。
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