Department of Materials, Department of Bioengineering, and Institute of Biomedical Engineering, Imperial College London, Exhibition Road, London, SW7 2AZ, UK.
Department of Physics, Chalmers University of Technology, 41296, Göteborg, Sweden.
Chembiochem. 2018 Mar 2;19(5):434-438. doi: 10.1002/cbic.201700592. Epub 2018 Jan 15.
Membrane fusion is a process of fundamental importance in biological systems that involves highly selective recognition mechanisms for the trafficking of molecular and ionic cargos. Mimicking natural membrane fusion mechanisms for the purpose of biosensor development holds great potential for amplified detection because relatively few highly discriminating targets lead to fusion and an accompanied engagement of a large payload of signal-generating molecules. In this work, sequence-specific DNA-mediated liposome fusion is used for the highly selective detection of microRNA. The detection of miR-29a, a known flu biomarker, is demonstrated down to 18 nm within 30 min with high specificity by using a standard laboratory microplate reader. Furthermore, one order of magnitude improvement in the limit of detection is demonstrated by using a novel imaging technique combined with an intensity fluctuation analysis, which is coined two-color fluorescence correlation microscopy.
膜融合是生物系统中具有重要意义的过程,涉及分子和离子货物运输的高度选择性识别机制。为了生物传感器的发展而模拟自然膜融合机制具有很大的放大检测潜力,因为相对较少的高度区分靶标导致融合,并伴随着大量信号生成分子的参与。在这项工作中,使用序列特异性 DNA 介导的脂质体融合来高度选择性地检测 microRNA。通过使用标准实验室微孔板读取器,在 30 分钟内以高特异性检测到 miR-29a(一种已知的流感生物标志物),检测下限低至 18nm。此外,通过使用结合强度波动分析的新型成像技术(称为双色荧光相关显微镜),检测下限提高了一个数量级。