Polymerase/Nicking Enzyme Powered Dual-template Multi-cycle G-Triplex Machine for HIV-1 Determination

作者全名："Yi, Gang; Duan, Qiuyue; Yan, Qi; Huang, Yuqi; Zhang, Wenxiu; Zhao, Shuhui"

作者地址："[Yi, Gang; Duan, Qiuyue; Yan, Qi; Huang, Yuqi; Zhang, Wenxiu; Zhao, Shuhui] Chongqing Med Univ, Dept Lab Med, Minist Educ China, Key Lab Clin Lab Diagnost, Chongqing 400016, Peoples R China"

通信作者："Yi, G (corresponding author), Chongqing Med Univ, Dept Lab Med, Minist Educ China, Key Lab Clin Lab Diagnost, Chongqing 400016, Peoples R China."

来源：ANALYTICAL SCIENCES

ESI学科分类：CHEMISTRY

WOS号：WOS:000681706100005

JCR分区：Q4

影响因子：1.6

年份：2021

卷号：37

期号：8

开始页：1087

结束页：1093

文献类型：Article

关键词：DNA machine; multi-cycle amplification; fluorescence; HIV; trace biomarker analysis

摘要："We proposed a dual-template, multi-cycle DNA nanomachine driven by polymerase and a nicking enzyme with high efficiency. The reaction system simply consists of two templates (Ti, T2) and two enzymes (KF polymerase, Nb.BbvCI). The two templates are similar in structure (X-X'-Y, Y-Y'-C) with a primer recognition region, a primer analogue generation region, an output region (3'-5'), and two nicking sites. The output strand of T1 is the primer of T2, and the G-rich fragment (G3) is designed as the final product. In the presence of HIV-1, numerous G3 were generated through the multi-cycle amplification strategy and formed a G-triplex/ThT complex after the addition of thioflavin T (ThT), which greatly enhanced the fluorescence intensity as a 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-cycle G-triplex machine, and benefiting from the high efficiency amplification strategy, the enzymatic reaction can be completed within 45 min and followed by fluorescence measurements. In addition, the analysis of other targets can be achieved by replacing the template sequence. Thus, there is a certain application potential of this strategy for trace biomarker analysis."

基金机构：Science and Technology Research Program of Chongqing Yuzhong District Science and Technology Commission [20180127]

基金资助正文：Thanks for the support of the Science and Technology Research Program of Chongqing Yuzhong District Science and Technology Commission (Grant No. 20180127).