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基于上转换磷的定量免疫层析分析检测 spp. 和孢子的校准。

Calibration of an Upconverting Phosphor-Based Quantitative Immunochromatographic Assay for Detecting spp., and Spores.

机构信息

State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China.

Beijing Key Laboratory of POCT for Bioemergency and Clinic, Beijing, China.

出版信息

Front Cell Infect Microbiol. 2020 Apr 24;10:147. doi: 10.3389/fcimb.2020.00147. eCollection 2020.

Abstract

spp., and are pathogens that can cause infectious zoonotic diseases with high mortality rates. An upconverting phosphor-based quantitative immunochromatographic (UPT-LF) assay, a point-of-care testing method suitable for resource-limited areas, was calibrated to quantitatively detect pathogenic bacteria. The bacterial purity or activity were ensured via staining methods and growth curves, respectively. Growth assays showed that the classic plate-counting method underestimated bacterial numbers compared with the bacterial counting method recommended by the reference material of the National Institutes for Food and Drug Control, China. The detection results of the UPT-LF assay differed significantly between the bacterial cultures in liquid and solid media and between different strains. Accelerated stability assessments and freeze-thaw experiments showed that the stability of the corresponding antigens played an important role in calibrating the UPT-LF assay. In this study, a new calibration system was developed for quantitative immunochromatography for detecting pathogenic bacteria. The results demonstrated the necessity of calibration for standardizing point-of-care testing methods.

摘要

spp. 和 是病原体,可引起具有高死亡率的传染性人畜共患病。上转换荧光粉定量免疫层析(UPT-LF)检测法是一种适合资源有限地区的即时检测方法,经校准后可定量检测病原菌。细菌纯度或活性分别通过染色方法和生长曲线来保证。生长实验表明,与中国食品药品检定研究院参考物质推荐的细菌计数方法相比,经典平板计数法低估了细菌数量。UPT-LF 检测法在液体和固体培养基中的细菌培养物以及不同菌株之间的检测结果差异显著。加速稳定性评估和冻融实验表明,相应抗原的稳定性在 UPT-LF 检测法的校准中起着重要作用。在本研究中,为检测病原菌的定量免疫层析开发了一种新的校准系统。结果表明,标准化即时检测方法需要进行校准。

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