Khreich Nathalie, Lamourette Patricia, Boutal Hervé, Devilliers Karine, Créminon Christophe, Volland Hervé
CEA, iBiTecS, Service de Pharmacologie et d'Immunoanalyse, Laboratoire d'Etudes et de Recherche en Immunoanalyse, F- 91191 Gif sur Yvette, France.
Anal Biochem. 2008 Jun 15;377(2):182-8. doi: 10.1016/j.ab.2008.02.032. Epub 2008 Mar 18.
Staphylococcus enterotoxin B (SEB) is one of several toxins produced by the gram positive bacterium Staphylococcus aureus. SEB is a major cause of food poisoning and represents a significant biological threat with regard to bioterrorism. A rapid, sensitive, and specific method is required to monitor food and water in cases of both natural and intentional contamination by this toxin. This report presents an improved immunochromatographic test (ICT) using immunoliposomes as label for the detection of SEB. For the first time in an ICT, the signal generated by the sulforhodamine B encapsulated into immunoliposomes was measured by fluorescence, allowing a 15-fold increase in sensitivity compared with that for visual detection of colored labels. The ICT was completed within 30 min, providing a limit of detection close to 20 pg/ml in buffer and showing no cross-reactivity with the other major toxin of the bacterium, Staphylococcus enterotoxin A. This sensitivity was retained when analyzing SEB spiked in various alimentary matrices, mimicking contaminated foods. Due to the use of fluorescent immunoliposomes as label, the present assay offers the inherent simplicity and speed of a dipstick assay while providing detection of low levels of SEB in real samples.
葡萄球菌肠毒素B(SEB)是革兰氏阳性菌金黄色葡萄球菌产生的几种毒素之一。SEB是食物中毒的主要原因,在生物恐怖主义方面构成重大生物威胁。需要一种快速、灵敏且特异的方法来监测食物和水是否受到这种毒素的自然污染或人为污染。本报告介绍了一种改进的免疫层析试验(ICT),该试验使用免疫脂质体作为标记物来检测SEB。在免疫层析试验中,首次通过荧光测量封装在免疫脂质体中的磺基罗丹明B产生的信号,与目视检测有色标记相比,灵敏度提高了15倍。该免疫层析试验在30分钟内完成,在缓冲液中的检测限接近20 pg/ml,并且与该细菌的另一种主要毒素葡萄球菌肠毒素A没有交叉反应。在分析添加到各种食品基质中的SEB(模拟受污染食品)时,这种灵敏度得以保持。由于使用荧光免疫脂质体作为标记物,本检测方法具有试纸条检测固有的简便性和速度,同时能够检测实际样品中的低水平SEB。