论文标题
聚合酶/昵称酶供电的双动型多循环G-Triplex机器用于HIV-1测定
Polymerase/nicking enzyme powered dual-template multi-cycled G-triplex machine for HIV-1 determination
论文作者
论文摘要
我们提出了一种由聚合酶刻痕酶高效驱动的双板多伴侣DNA纳米机械。反应系统仅由两个模板（T1，T2）和两个酶（KF聚合酶，NB.BBVCI）组成。这两个模板在结构（X-X-Y，Y-Y-C）上相似：底漆识别区域，引物模拟产生区域，输出区域（3至5），并且每个两个区域之间都有一个缺口位点。 T1的输出是T2和富含G的片段（G3）的底漆，被设计为最终产品。在存在HIV-1的情况下，由于多循环放大策略，产生了许多G3，并在添加硫非类黄素T（THT）后形成G-Triplex Tht复合物，从而在无标记的传感策略中大大增强了荧光强度为信号报道的荧光强度。 HIV-1基因检测的动态响应范围为50 fm-2 nm，可以通过这款多循环的G-TRIPLEX机器实现，并受益于高效率扩增策略，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对其他目标进行分析。因此，在此策略中有一定的痕量生物标志物分析的应用潜力。
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.
