Clinical Medicine Research Center, Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, PR China; Guangdong Engineering and Technology Research Center for Rapid Diagnostic Biosensors, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, PR China.
Clinical Medicine Research Center, Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, PR China; Department of Laboratory Medicine, People's Hospital of Shenzhen Baoan District, Shenzhen, 518101, PR China.
Anal Chim Acta. 2022 May 29;1209:339006. doi: 10.1016/j.aca.2021.339006. Epub 2021 Aug 30.
Intracellular substance analysis is critical for understanding cellular physiological mechanisms and predicting disease progression. Isothermal amplification technologies have been raised to accurately detect intracellular substances due to their low abundance, which is significant for the mechanism analysis and clinical application. However, traditional isothermal method still needs to cell destruction and extraction, resulting in fluctuant results. Moreover, it only works on dead cells. Therefore, non-destructive analysis based on isothermal amplification deserves to be studied, which directly reveals the content and position of relevant molecules. In recent years, metastable DNA hairpins-driven isothermal amplification (Mh-IA) blazes a trail for analysis in living cells. This review tracks the recent advances of Mh-IA strategy in living cell detection and highlights the potential challenges regarding this field, aiming to improve in vivo isothermal amplification. Also, challenges and prospects of Mh-IA for in situ and intracellular analysis are considered.
细胞内物质分析对于理解细胞生理机制和预测疾病进展至关重要。由于细胞内物质丰度低，等温扩增技术被提出以准确检测细胞内物质，这对于机制分析和临床应用具有重要意义。然而，传统的等温方法仍然需要细胞破坏和提取，导致结果不稳定。此外，它仅适用于死细胞。因此，基于等温扩增的非破坏性分析值得研究，它可以直接揭示相关分子的含量和位置。近年来，基于亚稳态 DNA 发夹驱动的等温扩增（Mh-IA）为活细胞分析开辟了道路。本综述跟踪了 Mh-IA 策略在活细胞检测中的最新进展，并强调了该领域的潜在挑战，旨在改善体内等温扩增。同时，还考虑了 Mh-IA 用于原位和细胞内分析的挑战和前景。
