Zhang Qingquan, Li Jiajia, Su Yuting, Pan Xiaoyan, Gai Hongwei
Jiangsu Key Laboratory of Green Synthesis for Functional Materials, School of Chemistry and Materials Science, Jiangsu Normal University Xuzhou Jiangsu 221116 China
Department of Laboratory Medicine, The Second Affiliated Hospital, Zhejiang University School of Medicine Hangzhou 310009 China.
RSC Adv. 2021 Aug 13;11(44):27541-27546. doi: 10.1039/d1ra04360a. eCollection 2021 Aug 9.
The relatively low sensitivity is the limitation of ELISA in early screening of diseases. Various signal enhancing methods are developed to increase detection sensitivity, but most of them require professional operation and are not compatible with commercial reagent kits or automatic analysis instruments, limiting their application in clinical testing. Here, we present a contactless and user-friendly ball-lens assisted fluorescence enhancing (BFE) method to improve immunoassay sensitivity. The BFE effect is observed in experiment and demonstrated by the simulation. Based on the BFE effect, the detection sensitivity of the immunoassay is improved about 3.6-fold and the limit of detection is lowered by more than 3-fold using commercial reagents and standard ELISA processes. In addition, the BFE effect causes no damage to the linear dependence, specificity and accuracy of clinical plasma measurement. The established method shows a high compatibility with automatic microfluidic immunoassay systems and other signal enhancing techniques, displaying great potential in multi-technique coupling assays and clinical applications.
相对较低的灵敏度是酶联免疫吸附测定法（ELISA）在疾病早期筛查中的局限性。人们开发了各种信号增强方法来提高检测灵敏度，但其中大多数方法需要专业操作，并且与商业试剂盒或自动分析仪器不兼容，限制了它们在临床检测中的应用。在此，我们提出一种非接触式且用户友好的球透镜辅助荧光增强（BFE）方法，以提高免疫测定的灵敏度。在实验中观察到了BFE效应，并通过模拟进行了验证。基于BFE效应，使用商业试剂和标准ELISA流程，免疫测定的检测灵敏度提高了约3.6倍，检测限降低了3倍以上。此外，BFE效应不会损害临床血浆测量的线性相关性、特异性和准确性。所建立的方法与自动微流控免疫测定系统和其他信号增强技术具有高度兼容性，在多技术耦合测定和临床应用中显示出巨大潜力。
