Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology , Wuhan University , Wuhan 430072 , P. R. China.
ACS Appl Mater Interfaces. 2019 Feb 13;11(6):5762-5770. doi: 10.1021/acsami.8b18898. Epub 2019 Feb 5.
The fluorescence method has made great progress in the construction of sensitive sensors but the background fluorescence of the matrix and photobleaching limit its broad application in clinical diagnosis. Here, we propose a digital single virus immunoassay for multiplex virus detection by using fluorescent magnetic multifunctional nanospheres as both capture carriers and signal labels. The superparamagnetism and strong magnetic response ability of nanospheres can realize efficient capture and separation of targets without sample pretreatment. Due to their distinguishable fluorescence imaging and photostability, the nanospheres enable single-particle counting for ultrasensitive multiplexed detection. Furthermore, the integration of digital analysis provided a reliable quantitative strategy for the detection of rare targets. Based on multifunctional nanospheres and digital analysis, a digital single virus immunoassay was proposed for simultaneous detection of H9N2, H1N1, and H7N9 avian influenza virus without complex signal amplification, whose detection limits were 0.02 pg/mL. Owing to its good specificity and anti-interference ability, the method showed great potential in single biomolecules, multiplexed detection, and early diagnosis of diseases.
荧光法在构建敏感传感器方面取得了很大进展，但基质的背景荧光和光漂白限制了其在临床诊断中的广泛应用。在这里，我们提出了一种数字单病毒免疫分析方法，用于通过使用荧光磁性多功能纳米球作为捕获载体和信号标记物来进行多重病毒检测。纳米球的超顺磁性和强磁响应能力可以实现对目标物的高效捕获和分离，而无需样品预处理。由于其可区分的荧光成像和光稳定性，纳米球能够进行单粒子计数，实现超灵敏的多重检测。此外，数字分析的集成提供了一种可靠的定量策略，用于检测稀有目标物。基于多功能纳米球和数字分析，提出了一种数字单病毒免疫分析方法，用于同时检测 H9N2、H1N1 和 H7N9 禽流感病毒，无需复杂的信号放大，其检测限为 0.02 pg/mL。由于其良好的特异性和抗干扰能力，该方法在单分子、多重检测和疾病的早期诊断方面具有很大的潜力。
