Magnetic Immunoassay for Detection of Staphylococcal Toxins in Complex Media
Magnetic Immunoassay for Detection of Staphylococcal Toxins in Complex Media
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DOI：
10.1021/ac303075b
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发表时间：
2013-01-15
影响因子：
7.4
通讯作者：
Nikitin, Petr I.
中科院分区：
文献类型：
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作者：
Orlov, Alexey V.;Khodakova, Julia A.;Nikitin, Petr I.
利用磁性纳米颗粒的非线性磁化高灵敏配准方法，建立了一种新型的夹心式免疫分析方法，用于检测几乎任何体积的复杂介质中的葡萄球菌毒素，随着样品体积的增加，灵敏度不断提高。信号是从用作固相的不透明3D纤维结构的整个体积中读出的，这提供了大的反应表面、快速的试剂混合以及在分析过程中直接的抗原免疫过滤。该方法在近30年的大范围内呈现出近乎线性的剂量-反应曲线，同时在不需要样品制备的情况下检测纯牛奶中的葡萄球菌肠毒素A(SEA)和中毒性休克综合征毒素(TSST)。对于30毫升的样品，在2小时格式下，TSST和SEA的检出限分别为4和10 pg/mL。第二份，25分钟格式，显示了150亩L样品中相同毒素的检出限分别为0.1%和0.3%。所开发的免疫分析方法可应用于食品安全控制、体外诊断和兽医领域的各种研究，从现场快速检测到高度敏感的实验室检测。
Method of highly sensitive registration of magnetic nanoparticles by their nonlinear magnetization is used in a novel sandwich-type immunoassay for detection of staphylococcal toxins in complex media of virtually any volume, with increasing sensitivity at higher sample volume. The signal is read out from the entire volume of a nontransparent 3D fiber structure employed as a solid phase, which provides large reaction surface, quick reagent mixing, as well as antigen immunofiltration directly in the course of the assay. The method has demonstrated near-linear dose-response curves within a wide range of similar to 3 decades, while detection of staphylococcal enterotoxin A (SEA) and toxic shock syndrome toxin (TSST) in neat milk without sample preparation. The limits of detection (LOD) as low as 4 and 10 pg/mL for TSST and SEA, respectively, were obtained in 2-h format using 30-mL samples. The second, 25-min format, showed the LOD of 0.1 and 0.3 ng/mL for the same toxins in a 150 mu L sample. The developed immunoassay can be applied in food safety control, in vitro diagnostics, and veterinary for a variety of research from express tests in the field to highly sensitive laboratory tests.