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镧系元素标记免疫层析条用于快速检测斯氏泛菌亚种。

Lanthanide-labeled immunochromatographic strips for the rapid detection of Pantoea stewartii subsp. stewartii.

机构信息

Chinese Academy of Inspection and Quarantine, Beijing 100123, China.

出版信息

Biosens Bioelectron. 2014 Jan 15;51:29-35. doi: 10.1016/j.bios.2013.06.065. Epub 2013 Jul 19.

DOI:10.1016/j.bios.2013.06.065
PMID:23928093
Abstract

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 colorimetric 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 10(4)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.

摘要

侧向流动免疫分析用于商业妊娠检测,是一种被认可的即时检测技术。最广泛使用的侧向流动免疫层析条格式使用金纳米颗粒进行比色检测。然而,这种方法常常受到定量分辨力差和分析灵敏度低的限制。为了解决这些限制,已将镧系螯合物负载的硅纳米颗粒用作荧光标记物。荧光纳米颗粒可以很容易地与抗体结合,以葡聚糖作为连接物。这里描述的条带读取器是基于在条带上进行的夹心免疫反应,使用镧系标记的抗体作为荧光读出的信号载体。条带读取器用作定量测试系统。在这项工作中,以 Pantoea stewartii subsp. stewartii(Pss)作为模型分析物来演示条带读取器的用途。在最佳条件下,检测限确定为 10(3)cfu/mL。定量限计算为 10(4)cfu/mL。Pss 的检测限比胶体金标记条或 ELISA 显示的低 100 倍。与其他九种菌株没有观察到交叉反应,表明 Pss 条具有良好的特异性。该条在重复测试中表现出良好的稳定性。荧光免疫层析条的测试易于操作、快速且灵敏。使用荧光条和条带读取器的方法有可能成为定量细菌的有力工具。

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