Gold nanoparticle-based enhanced chemiluminescence immunosensor for detection of Staphylococcal Enterotoxin B (SEB) in food.
Gold nanoparticle-based enhanced chemiluminescence immunosensor for detection of Staphylococcal Enterotoxin B (SEB) in food.
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DOI：
10.1016/j.ijfoodmicro.2009.05.029
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发表时间：
2009-08-15
影响因子：
5.4
通讯作者：
Rasooly A
中科院分区：
文献类型：
--
作者：
Yang M;Kostov Y;Bruck HA;Rasooly A
葡萄球菌肠毒素（Staphylococcal enterotoxins，SE）是引起食源性疾病的主要原因，因此需要灵敏度<1 ng/ml的检测方法。金纳米颗粒的表面积、几何和物理性质使其非常适合在测定中增强与生物分子的相互作用。利用金纳米粒子的免疫检测特性，我们研制了一种基于金纳米粒子的增强化学发光（ECL）免疫传感器，用于检测食品中的葡萄球菌肠毒素B（SE B）。通过物理吸附将抗SEB一抗固定到金纳米颗粒表面上，然后将抗体-金纳米颗粒混合物固定到聚碳酸酯表面上。SEB通过在聚碳酸酯表面上的“ELISA型”ELISA测定用二抗和ECL检测来检测。使用基于冷却电荷耦合器件（CCD）传感器或读板器的即时检测器读取ECL的信号。该系统用于测试缓冲液和各种食物（蘑菇，西红柿和婴儿食品肉类）中的SEB。发现检测限为~0.01 ng/mL，比传统ELISA灵敏度高~10倍。金纳米颗粒由于其物理吸附机制而相对容易用于抗体固定;不需要其他试剂进行固定。使用我们的简单和廉价的检测器结合金纳米粒子的ECL方法在这里描述的是适用于简化和提高任何免疫学测定和即时诊断的灵敏度。
Staphylococcal enterotoxins (SEs) are major cause of foodborne diseases, so sensitive detection (<1 ng/ml) methods are needed for SE detection in food. The surface area, geometric and physical properties of gold nanoparticles make them well-suited for enhancing interactions with biological molecules in assays. To take advantage of the properties of gold nanoparticles for immunodetection, we have developed a gold nanoparticle-based enhanced chemiluminescence (ECL) immunosensor for detection of Staphylococcal Enterotoxin B (SEB) in food. Anti-SEB primary antibodies were immobilized onto a gold nanoparticle surface through physical adsorption and then the antibody–gold nanoparticle mixture was immobilized onto a polycarbonate surface. SEB was detected by a “sandwich-type” ELISA assay on the polycarbonate surface with a secondary antibody and ECL detection. The signal from ECL was read using a point-of-care detector based on a cooled charge-coupled device (CCD) sensor or a plate reader. The system was used to test for SEB in buffer and various foods (mushrooms, tomatoes, and baby food meat). The limit of detection was found to be ~0.01 ng/mL, which is ~10 times more sensitive than traditional ELISA. The gold nanoparticles were relatively easy to use for antibody immobilization because of their physical adsorption mechanism; no other reagents were required for immobilization. The use of our simple and inexpensive detector combined with the gold nanoparticle-based ECL method described here is adaptable to simplify and increase sensitivity of any immunological assay and for point-of-care diagnostics.
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