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Development and evaluation of an up-converting phosphor technology-based lateral flowassay for rapid detection of Francisella tularensis

机译:基于上转换荧光粉技术的侧向流的开发和评估法快速检测土拉弗朗西斯菌

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摘要

Francisella tularensis is a potential biowarfare/bioterrorism agent and zoonotic pathogen that causes tularemia; thus, surveillance of F. tularensis and first-level emergency response using point-of-care testing (POCT) are essential. The UPT-LF POCT assay was established to quantitatively detect F. tularensis within 15 min, and the sensitivity of the assay was 104 CFU · mL−1 (100 CFU/test). The linear quantitative range covered five orders of magnitude, and the coefficients of variation were less than 10%. Except Shigella dysenteriae, UPT-LF showed excellent specificity to four strains that are also potential biowarfare/bioterrorism agents and 13 food-borne pathogenic strains. Samples with pH 2–13, high ion strengths (≥2 mol · L−1 solution of KCl and NaCl), high viscosities (≤50 mg · mL−1 or ≥20% glycerol), and high concentrations of biomacromolecules (≥400 mg · mL−1 bovine serum albumin or≥80 mg · mL−1casein) showed little influence on the assay. For practical utilization, thetolerance limits for seven powders and eight viscera were determined, and operationerrors of liquid measurement demonstrated a minor influence on the strip. Ftu-UPT-LFis a candidate POCT method because of its excellent sensitivity, specificity, andstability in complex samples, as well as low operation error.
机译:图拉弗朗西斯菌(Francisella tularensis)是潜在的生物战/生物恐怖分子和引起图拉菌血症的人畜共患病原体。因此,使用即时检验(POCT)监视土拉弗朗西斯菌和一级紧急响应至关重要。建立了UPT-LF POCT检测方法,以在15 min内定量检测土拉参。检测的灵敏度为10 4 CFU·mL -1 (100 CFU /测试)。线性定量范围涵盖五个数量级,变异系数小于10%。除痢疾志贺氏菌外,UPT-LF对4种也是潜在的生物战/生物恐怖主义制剂的菌株和13种食源性致病菌株显示出优异的特异性。 pH 2–13,高离子强度(KCl和NaCl的≥2≥mol·L -1 溶液),高粘度(≤50mg·mL -1 或≥20%甘油)和高浓度的生物大分子(≥400mg·mL −1 牛血清白蛋白或≥80mg·mL -1 酪蛋白)对测定几乎没有影响。为了实际使用,确定了七个粉末和八个内脏的公差极限,并进行了操作液体测量的误差显示出对带材的轻微影响。 Ftu-UPT-LF由于其出色的灵敏度,特异性和复杂样品的稳定性好,操作误差低。

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