Department of Chemical and Biological Engineering, Drexel University, Philadelphia, PA 19104, USA.
Analyst. 2014 Apr 7;139(7):1576-88. doi: 10.1039/c3an01677c.
The current state of biosensor-based techniques for amplification-free microRNA (miRNA) detection is critically reviewed. Comparison with non-sensor and amplification-based molecular techniques (MTs), such as polymerase-based methods, is made in terms of transduction mechanism, associated protocol, and sensitivity. Challenges associated with miRNA hybridization thermodynamics which affect assay selectivity and amplification bias are briefly discussed. Electrochemical, electromechanical, and optical classes of miRNA biosensors are reviewed in terms of transduction mechanism, limit of detection (LOD), time-to-results (TTR), multiplexing potential, and measurement robustness. Current trends suggest that biosensor-based techniques (BTs) for miRNA assay will complement MTs due to the advantages of amplification-free detection, LOD being femtomolar (fM)-attomolar (aM), short TTR, multiplexing capability, and minimal sample preparation requirement. Areas of future importance in miRNA BT development are presented which include focus on achieving high measurement confidence and multiplexing capabilities.
本文批判性地回顾了基于生物传感器的无扩增微 RNA（miRNA）检测技术的现状。从转导机制、相关方案和灵敏度等方面，将其与非传感器和基于扩增的分子技术（MTs），如聚合酶方法进行了比较。简要讨论了 miRNA 杂交热力学中影响分析物选择性和扩增偏差的挑战。根据转导机制、检测限（LOD）、结果获得时间（TTR）、多重检测潜力和测量稳健性，对电化学、机电和光学 miRNA 生物传感器进行了综述。目前的趋势表明，由于无扩增检测的优势，基于生物传感器的技术（BTs）将补充 MTs，其检测限为飞摩尔（fM）-皮摩尔（pM），获得结果的时间短（TTR），具有多重检测能力，且对样品制备的要求很少。提出了 miRNA BT 未来发展的重要领域，包括关注提高测量置信度和多重检测能力。
