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通过优化参数开发用于检测食品病原体沙门氏菌的快速试纸条检测方法。

Development of rapid dipstick assay for food pathogens, Salmonella, by optimized parameters.

作者信息

Çam Dilek, Öktem Hüseyin Avni

机构信息

1Department of Biological Sciences, Middle East Technical University, 06800 Ankara, Turkey.

2Department of Biology, Çankırı Karatekin University, 18100 Çankırı, Turkey.

出版信息

J Food Sci Technol. 2019 Jan;56(1):140-148. doi: 10.1007/s13197-018-3467-5. Epub 2018 Oct 25.

Abstract

Salmonella is among the very important pathogens threating the human and animal health. Rapid and easy detection of these pathogens is crucial. In this context, antibody (Ab) based lateral flow assays (LFAs) which are simple immunochromatographic point of care test kits were developed by gold nanoparticles (GNPs) as labelling agent for Salmonella detection. For that purpose some critical parameters such as reagent concentrations on the capture zones, conjugate concentrations and ideal membrane type needed for LFAs for whole cell detection were tested for naked eye analysis. Therefore, prepared LFAs were applied to the live and heat inactivated cells when they were used alone or included in different bacterial mixtures. Among the test platforms, membrane 180 (M180) was found as an ideal membrane and 36 nm GNPs showed highly good labelling in the developed LFAs. Diluted conjugates and low concentrations of reagents affected the test signal negatively. Salmonella was detected in different bacterial mixtures, selectively in 4-5 min. The best recognized species by used Ab were and . 5 × 10 cells were also determined as a limit of detection of this study with mentioned parameters.

摘要

沙门氏菌是威胁人类和动物健康的非常重要的病原体之一。快速简便地检测这些病原体至关重要。在此背景下,基于抗体(Ab)的侧向流动分析(LFA),即简单的免疫层析即时检测试剂盒,采用金纳米颗粒(GNP)作为标记剂用于沙门氏菌检测而得以开发。为此,针对全细胞检测的LFA所需的一些关键参数,如捕获区的试剂浓度、偶联物浓度和理想的膜类型,进行了肉眼分析测试。因此,制备好的LFA在单独使用或包含在不同细菌混合物中时,应用于活细胞和热灭活细胞。在测试平台中,膜180(M180)被发现是理想的膜,并且36纳米的GNP在开发的LFA中显示出非常好的标记效果。稀释的偶联物和低浓度的试剂对测试信号有负面影响。在不同细菌混合物中能选择性地在4 - 5分钟内检测到沙门氏菌。所用抗体最易识别的菌种是 和 。在上述参数下,本研究还确定5×10 个细胞为检测限。

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