Lanthanide-labeled immunochromatographic strips for the rapid detection of Pantoea stewartii subsp stewartii
Lanthanide-labeled immunochromatographic strips for the rapid detection of Pantoea stewartii subsp stewartii
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DOI：
10.1016/j.bios.2013.06.065
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发表时间：
2014-01-15
影响因子：
12.6
通讯作者：
Xue, Qiang
中科院分区：
文献类型：
--
作者：
Zhang, Fan;Zou, Mingqiang;Xue, Qiang
侧流免疫测定用于商业妊娠检测，是一种公认的即时检测技术。最广泛使用的侧流免疫层析试纸条使用金纳米粒子进行量热检测。然而，该方法常常存在定量辨别力差和分析灵敏度低的问题。为了解决这些限制，负载有镧系元素螯合物的二氧化硅纳米粒子已被用作荧光标记。荧光纳米粒子可以很容易地与抗体结合，以葡聚糖作为连接体。这里描述的条带读取器基于在条带上进行的夹心免疫反应，使用镧系元素标记的抗体作为荧光读数的信号载体。试纸条读取器用作定量测试系统。在这项工作中，Pantoea Steartii subsp。 Steartii (Pss) 被用作模型分析物来演示试纸条读取器的使用。在最佳条件下，检测限确定为10(3) cfu/mL。计算得出的定量限为 104 cfu/mL。 Pss 的检测限比胶体金标记试纸条或 ELISA 的检测限低 100 倍。未观察到与其他 9 个菌株的交叉反应，表明 Pss 试纸条具有良好的特异性。该条带在重复测试中表现出良好的稳定性。使用荧光免疫层析试纸条进行的测试易于执行、快速且灵敏。使用荧光条和条阅读器的方法有可能成为细菌定量的强大工具。 (c) 2013 Elsevier B.V. 保留所有权利。
The lateral flow immunoassay is used in commercial pregnancy detection, and is an accepted point-of-care testing technique. The most widely used format for lateral flow immunochromatographic strips uses gold nanoparticles for calorimetric detection. However, this method often suffers from poor quantitative discrimination and low analytical sensitivity. To address these limitations, lanthanide chelate-loaded silica nanoparticles have been used as fluorescent labels. The fluorescent nanoparticles can easily bind to antibodies, with dextran as a linker. The strip reader described here was based on a sandwich immunoreaction performed on a strip, using lanthanide-labeled antibodies that served as signal vehicles for the fluorescent readout. The strip reader was used as a quantitative test system. In this work, Pantoea stewartii subsp. stewartii (Pss) was used as a model analyte to demonstrate the use of the strip reader. Under optimal conditions, the detection limit was determined as 10(3) cfu/mL. The quantification limit was calculated to be 104 cfu/mL. The detection limit for Pss was 100 times lower than those displayed by colloidal gold-labeled strips or ELISAs. No cross-reactions were observed with the other nine strains, indicating the good specificity of the Pss strip. This strip showed good stability in repeated tests. The tests using the fluorescence immunochromatographic strip were easy to perform, rapid, and sensitive. Methods using fluorescence strips and a strip reader have the potential to be a powerful tool for the quantification of bacteria. (c) 2013 Elsevier B.V. All rights reserved.