A low-cost, high-performance system for fluorescence lateral flow assays.
A low-cost, high-performance system for fluorescence lateral flow assays.
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用于荧光横向流量测定的低成本高性能系统。
DOI：
10.3390/bios3040360
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发表时间：
2013-10-21
期刊：
影响因子：
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通讯作者：
Oldham MF
中科院分区：
文献类型：
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作者：
Lee LG;Nordman ES;Johnson MD;Oldham MF
我们展示了一个荧光侧向流动系统，具有良好的灵敏度和宽的动态范围。照明系统利用LED、塑料透镜以及塑料和有色玻璃滤光器用于激发和发射光。图像是在iPhone 4上收集的。几种具有长斯托克斯位移的荧光染料在侧向流中的信号和非特异性结合进行了评价。发现信号与非特异性结合的比值范围很广，从R-藻红蛋白（R-PE）的50到亮紫605的0.15。R-PE的长斯托克斯位移允许使用廉价的塑料滤光片而不是昂贵的干涉滤光片来阻挡LED光。以生物素化牛血清白蛋白（BSA）为分析物，在侧流中直接比较了R-PE荧光检测法和胶体金吸光度检测法。荧光在0.4- 4，000 ng/mL范围内提供线性数据，信号变化为1，000倍，而胶体金在16- 4，000 ng/mL范围内提供非线性数据，信号变化为10倍。使用人绒毛膜促性腺激素（hCG）作为分析物的比较显示了类似的优势，在荧光系统。我们相信，我们的廉价而高性能的平台将是有用的，提供定量和灵敏的检测在一个点的护理设置。
We demonstrate a fluorescence lateral flow system that has excellent sensitivity and wide dynamic range. The illumination system utilizes an LED, plastic lenses and plastic and colored glass filters for the excitation and emission light. Images are collected on an iPhone 4. Several fluorescent dyes with long Stokes shifts were evaluated for their signal and nonspecific binding in lateral flow. A wide range of values for the ratio of signal to nonspecific binding was found, from 50 for R-phycoerythrin (R-PE) to 0.15 for Brilliant Violet 605. The long Stokes shift of R-PE allowed the use of inexpensive plastic filters rather than costly interference filters to block the LED light. Fluorescence detection with R-PE and absorbance detection with colloidal gold were directly compared in lateral flow using biotinylated bovine serum albumen (BSA) as the analyte. Fluorescence provided linear data over a range of 0.4–4,000 ng/mL with a 1,000-fold signal change while colloidal gold provided non-linear data over a range of 16–4,000 ng/mL with a 10-fold signal change. A comparison using human chorionic gonadotropin (hCG) as the analyte showed a similar advantage in the fluorescent system. We believe our inexpensive yet high-performance platform will be useful for providing quantitative and sensitive detection in a point-of-care setting.