Department of Nanoscience and Technology, Gachon University, 1342 Seongnam-daero, Sujeong-gu, Seongnam-si, Gyeonggi-do 13120, South Korea.
Department of BioNano Technology, Gachon University, 1342 Seongnam-daero, Sujeong-gu, Seongnam-si, Gyeonggi-do 13120, South Korea.
J Mater Chem B. 2024 Oct 9;12(39):10082-10092. doi: 10.1039/d4tb01623h.
In this investigation, the photophysical properties and interaction mechanisms of Janus green blue (JGB) dye with polyanions were systematically studied using spectroscopic techniques. The absorption spectral analysis revealed that JGB binds cooperatively to sodium alginate, leading to dye stacking along the polymer chain. The interaction of JGB dye with DNA was characterized by the emergence of a metachromatic peak at 564 nm, indicating the formation of dye aggregates. The analysis of absorption data reveals that JGB dye interacts with DNA at multiple binding sites, including at least one high-affinity site. The AutoDock Vina based blind docking approach was used to analyze the most probable binding location of JGB dye in DNA. By making use of the DNA-induced metachromasia, a colorimetric approach was developed for the visualization of loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR). The LAMP-colorimetric assay, targeting the gene, demonstrated a noticeable colour change with a detection limit of 1 pg μL. The practical applicability was validated by detecting in artificial urine. In addition to LAMP, we tested the JGB dye based colorimetric assay for applicability in PCR reactions. The colorimetric PCR assay using the metal-responsive transcription factor (MTF-1) gene achieved a detection limit as low as 0.1 pg μL. The study highlights the potential of DNA binding metachromic dye to significantly enhance colorimetric assays, offering a robust and sensitive tool for molecular diagnostics.
在这项研究中，我们使用光谱技术系统地研究了 Janus 绿蓝（JGB）染料与聚阴离子的光物理性质和相互作用机制。吸收光谱分析表明，JGB 与海藻酸钠协同结合，导致染料沿着聚合物链堆叠。JGB 染料与 DNA 的相互作用通过在 564nm 处出现变色峰来表征，表明形成了染料聚集体。吸收数据分析表明，JGB 染料在多个结合位点与 DNA 相互作用，至少包括一个高亲和力结合位点。使用 AutoDock Vina 基于盲目对接方法分析 JGB 染料在 DNA 中的最可能结合位置。利用 DNA 诱导的变色，开发了一种用于可视化环介导等温扩增（LAMP）和聚合酶链反应（PCR）的比色方法。针对 基因的 LAMP-比色测定法显示出明显的颜色变化，检测限为 1pg μL。通过检测人工尿液中的 验证了实际适用性。除了 LAMP，我们还测试了基于 JGB 染料的比色测定法在 PCR 反应中的适用性。使用金属反应转录因子（MTF-1）基因的比色 PCR 测定法可实现低至 0.1pg μL 的检测限。该研究强调了 DNA 结合变色染料显著增强比色测定法的潜力，为分子诊断提供了一种稳健而敏感的工具。
