Department of Applied Chemistry for Environment, Graduate School of Urban Environmental Sciences , Tokyo Metropolitan University , 1-1 Minamiohsawa, Hachioji , Tokyo 192-0397 , Japan.
School of Pharmacy , Fudan University , 826 Zhangheng Road , Shanghai 201203 , China.
Anal Chem. 2019 May 7;91(9):5685-5689. doi: 10.1021/acs.analchem.8b05668. Epub 2019 Apr 22.
The microwell plate/microtiter plate is among the most widely used tools in immune assays. In this paper, we report on a sensitive method for enhancing fluorescence emission detection by simply adding several droplets of an immiscible organic compound into the microwells before detection. To prove the concept, human IgA was determined on a microwell plate using this droplet enhanced fluorescence (DEF) detection method. An obvious enhancement in fluorescence was observed. The detection limit (LOD) was about 1/20 times and the sensitivity was 4 times greater than that without droplets. To prove the use of the method for disease diagnosis, the IgG of measles in human plasma was determined using the proposed DEF method. A LOD of around 1/5 times and a sensitivity of 4 times the DEF were easily achieved compared to ELISA with a conventional fluorescence detection.
微孔板/微量滴定板是免疫分析中应用最广泛的工具之一。在本文中，我们报告了一种简单的方法，通过在检测前向微孔中添加几滴不混溶的有机化合物，来增强荧光发射检测。为了验证这一概念，我们使用这种液滴增强荧光（DEF）检测方法在微孔板上测定了人 IgA。观察到荧光明显增强。检测限（LOD）约为无液滴时的 1/20 倍，灵敏度提高了 4 倍。为了证明该方法在疾病诊断中的应用，我们使用所提出的 DEF 方法测定了人血浆中麻疹的 IgG。与传统荧光检测的 ELISA 相比，该方法的 LOD 约为 1/5 倍，灵敏度提高了 4 倍。
