聚合酶/切口酶驱动的双模板多循环G-三链体仪测定HIV-1
Polymerase/nicking enzyme powered dual-template multi-cycled G-triplex machine for HIV-1 determination
摘要：
提出了一种聚合酶切口酶驱动的双模板多循环DNA纳米机器。反应系统简单地由两个模板（T1，T2）和两种酶（KF聚合酶，Nb.BbvCI）组成。两个模板在结构上相似（X-X-Y，Y-Y-C）：引物识别区、引物类似物产生区、输出区（3至5），并且在每两个区域之间存在切口位点。T1的输出产物为T2的引物，并设计了富含G的片段（G3）作为最终产物。在HIV-1的存在下，由于多循环扩增策略产生了大量的G3，并且在加入硫磺素T（ThT）后形成G-三链体ThT复合物，这大大增强了无标记传感策略中作为信号报告子的荧光强度。通过这种多循环的G-triplex机器可以实现50 fM-2 nM的HIV-1基因检测的动态响应范围，并且受益于高效的扩增策略，酶促反应可以在45分钟内完成，然后进行荧光测量。此外，可以通过替换模板序列来实现对其他靶标的分析。因此，在该策略中痕量生物标志物分析具有一定的应用潜力。
英文摘要：
We proposed a dual-template multi-cycled DNA nanomachine driven by polymerase nicking enzyme with high efficiency. The reaction system simply consists of two templates (T1, T2) and two enzymes (KF polymerase, Nb.BbvCI). The two templates are similar in structure (X-X-Y, Y-Y-C): primer recognition region, primer analogue generation region, output region (3 to 5), and there is a nicking site between each two regions. Output of T1 is the primer of T2 and G-rich fragment (G3) is designed as the final products. In the presence of HIV-1, numerous of G3 were generated owing to the multi-cycled amplification strategy and formed into G-triplex ThT complex after the addition of thioflavin T (ThT), which greatly enhanced the fluorescence intensity as signal reporter in the label-free sensing strategy. A dynamic response range of 50 fM-2 nM for HIV-1 gene detection can be achieved through this multi-cycled G-triplex machine, and benefit from the high efficiency amplification strategy, enzymatic reaction can be completed within 45 minutes followed by fluorescence measurement. In addition, analysis of other targets can be achieved by replacing the template sequence. Thus there is a certain application potential for trace biomarker analysis in this strategy.
机构、总结、中文摘要均由AI分析而来，可能存在错误，请以论文正文为准。
