Qin Chuanying, Hu Cong, Yu Aimin, Lai Guosong
Hubei Key Laboratory of Pollutant Analysis & Reuse Technology, College of Chemistry and Chemical Engineering, Hubei Normal University, Huangshi 435002, PR China.
Department of Chemistry and Biotechnology, Faculty of Science, Engineering and Technology, Swinburne University of Technology, Hawthorn, VIC 3122, Australia.
Analyst. 2021 Feb 22;146(4):1414-1420. doi: 10.1039/d0an02187c.
Herein, we report a Fe3O4@polydopamine (PDA) nanocomposite and exonuclease III (Exo III)-assisted homogeneous fluorescence biosensing method for ultrasensitive detection of kanamycin (Kana) antibiotic. A hairpin DNA containing the Kana-aptamer sequence (HP) was first designed for the highly specific biorecognition of the target analyte. Because of the aptamer biorecognition-induced structural change of HP and the highly effective catalyzed reaction of Exo III, a large amount of fluorophore labels were released from the designed fluorescence DNA probe. During the homogeneous reaction process, the Exo III-assisted dual recycling significantly amplified the fluorescence signal output. Moreover, the excessive probes were easily adsorbed and separated by the Fe3O4@PDA nanocomposite, which decreased the background signal and increased the signal-to-noise ratio. These strategies result in the excellent analytical performance of the method, including a very low detection limit of 0.023 pg mL-1 and a very wide linear range of six orders of magnitude. In addition, this method has convenient operation, excellent selectivity, repeatability and satisfactory reliability, and does not involve the design and utilization of complicated DNA sequences. Thus, it exhibits a promising prospect for practical applications.
在此,我们报道了一种用于超灵敏检测卡那霉素(Kana)抗生素的Fe3O4@聚多巴胺(PDA)纳米复合材料和外切核酸酶III(Exo III)辅助的均相荧光生物传感方法。首先设计了一种包含卡那霉素适配体序列的发夹DNA(HP),用于对目标分析物进行高特异性生物识别。由于适配体生物识别引起的HP结构变化以及Exo III的高效催化反应,大量荧光团标记从设计的荧光DNA探针中释放出来。在均相反应过程中,Exo III辅助的双重循环显著放大了荧光信号输出。此外,过量的探针很容易被Fe3O4@PDA纳米复合材料吸附和分离,从而降低了背景信号并提高了信噪比。这些策略使该方法具有出色的分析性能,包括极低的检测限0.023 pg mL-1和六个数量级的极宽线性范围。此外,该方法操作简便,选择性、重复性和可靠性良好,且不涉及复杂DNA序列的设计和应用。因此,它在实际应用中展现出广阔的前景。