基于水溶性氟硼二吡咯类共轭聚合物PBF可与长波长区(> 600 nm)荧光染料产生荧光共振能量转移(FRET),结合引物延伸反应,发展了一种均相检测miRNA的新方法.方法首先通过引物延伸反应在目标miRNA分子上引入荧光染料Cy5,形成miRNA-Cy5. 然后,PBF与miRNA-Cy5/DNA杂合体通过静电力结合并发生从PBF到Cy5的有效FRET,实现了基于阳离子共轭聚合物(CCP)的长波长区miRNA的均相检测,方法灵敏度高、特异性好,测定miR-221的线性为15~6 000 pmol/L,检出限(3σ)为8.4 pmol/L.方法拓展了CCP的应用,为基于CCP的生物传感和生化分析提供了新的均相检测平台.
Based on boron-dipyrromethene conjugated polymer PBF, which can generate fluorescence resonance energy transfer(FRET)with long-wavelength region(>600 nm)fluorescent dyes, a new homogeneous detection method for miRNA has been developed by combining with primer extension reaction. Firstly, the fluorescent dye Cy5 is introduced into the target miRNA by primer extension reaction to form miRNA-Cy5. Then, PBF and miRNA-Cy5/DNA hybrids are combined by electrostatic force and take place effective FRET from PBF to Cy5. The homogeneous detection of miRNA in long-wavelength based on cationic conjugated polymer(CCP)is realized, which has high sensitivity and specificity. The linear range for miR-221 is 15-6 000 pmol/L, and the detection limit(3σ)is 8.4 pmol/L. The method expands the applications of CCP and provides a new homogeneous detection platform for CCP-based biosensing and biochemical analysis.
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