A combined immunomagnetic separation and lateral flow method for a sensitive on-site detection of Bacillus anthracis spores - assessment in water and dairy products
A combined immunomagnetic separation and lateral flow method for a sensitive on-site detection of Bacillus anthracis spores - assessment in water and dairy products
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DOI：
10.1111/j.1472-765x.2008.02542.x
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发表时间：
2009-04-01
影响因子：
2.4
通讯作者：
Yitzhaki, S.
中科院分区：
文献类型：
--
作者：
Fisher, M.;Atiya-Nasagi, Y.;Yitzhaki, S.
结合免疫磁性分离（IMS）和侧流装置（LFD）检测，开发一种灵敏、快速的现场方法，能够浓缩和检测复杂样品中的炭疽芽孢杆菌孢子。数据显示，开发了一种优化的30 min IMS检测方法，B捕获率约为95%。来自不同乳制品的炭疽孢子（n = 38）。与典型乳植物群及部分密切相关的杆菌无交叉反应。为了能够将IMS捕获的孢子直接应用于LFD，在微波炉中使用95%（v/v）甲酰胺-10 mmol l（-1）EDTA从珠-孢子复合物中洗脱孢子30 s。分离的孢子在LFD上进行分析，使检测在10分钟内完成。与直接应用于LFD而不进行IMS浓缩步骤的样品相比，IMS-LFD组合方法（40分钟）的灵敏度提高了60倍。IMS-LFD方法是一个强大的平台，结合了浓缩和检测B的快速性、特异性和效率。IMS和LFD的结合提高了B的灵敏度和灵活性。炭疽孢子检测复杂的样品。这种方法可以扩展到各种基质中的其他毒素和微生物。
Combination of immunomagnetic separation (IMS) and lateral flow device (LFD) assays for the development of a sensitive, rapid, on-site methodology that enables concentration and detection of Bacillus anthracis spores in complex samples.The data presents the development of an optimized, 30 min, IMS assay, with about 95% capture of B. anthracis spores from different dairy products (n = 38). No cross reactivity was detected with typical milk flora and some closely related Bacilli. To enable direct application of the IMS captured spores on the LFD, spores were eluted from the bead-spore complex utilizing 95% (v/v) formamide-10 mmol l(-1) EDTA for 30 s in a microwave oven. Detached spores were analysed on LFD enabling detection within 10 min. The combined IMS-LFD methodology (40 min) demonstrates a 60-fold improvement in sensitivity, relative to samples that were applied directly on the LFD without the IMS concentrating step.The IMS-LFD method is a powerful platform, combining rapidity, specificity and efficiency for concentrating and detecting B. anthracis from water and milk contaminated samples.The combination of IMS and LFD enhances the sensitivity and flexibility of B. anthracis spore detection from complex samples. This method can potentially be extended to other toxins and micro-organisms in a variety of matrices.