Guangdong Provincial Key Laboratory of New Drug Screening and Guangzhou Key Laboratory of Drug Research for Emerging Virus Prevention and Treatment, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, PR China.
Department of Chemistry and Materials Engineering, Jiangsu Key Laboratory of Advanced Functional Materials, Changshu Institute of Technology, Changshu 215500, PR China.
Talanta. 2018 Apr 1;180:396-402. doi: 10.1016/j.talanta.2017.12.045. Epub 2017 Dec 15.
From a three-dimensional (3D) metal-organic framework (MOF) of {[Cu(Cmdcp)(phen)(HO)]·9HO} (1, HCmdcpBr = N-carboxymethyl-(3,5-dicarboxyl)pyridinium bromide, phen = phenanthroline), a sensitive and selective fluorescence sensor has been developed for the simultaneous detection of ebolavirus conserved RNA sequences and ebolavirus-encoded microRNA-like (miRNA-like) fragment. The results from molecular dynamics simulation confirmed that MOF 1 absorbs carboxyfluorescein (FAM)-tagged and 5(6)-carboxyrhodamine, triethylammonium salt (ROX)-tagged probe ss-DNA (probe DNA, P-DNA) by ππ stacking and hydrogen bonding, as well as additional electrostatic interactions to form a sensing platform of P-DNAs@1 with quenched FAM and ROX fluorescence. In the presence of targeted ebolavirus conserved RNA sequences or ebolavirus-encoded miRNA-like fragment, the fluorophore-labeled P-DNA hybridizes with the analyte to give a P-DNA@RNA duplex and released from MOF 1, triggering a fluorescence recovery. Simultaneous detection of two target RNAs has also been realized by single and synchronous fluorescence analysis. The formed sensing platform shows high sensitivity for ebolavirus conserved RNA sequences and ebolavirus-encoded miRNA-like fragment with detection limits at the picomolar level and high selectivity without cross-reaction between the two probes. MOF 1 thus shows the potential as an effective fluorescent sensing platform for the synchronous detection of two ebolavirus-related sequences, and offer improved diagnostic accuracy of Ebola virus disease.
从三维（3D）金属有机骨架（MOF）[{[Cu（Cmdcp）（phen）（HO）]·9HO}（1， HCmdcpBr = N-羧甲基-（3,5-二羧基）吡啶溴化物，phen = 菲咯啉）]，开发了一种用于同时检测埃博拉病毒保守 RNA 序列和埃博拉病毒编码的微 RNA 样（miRNA 样）片段的灵敏和选择性荧光传感器。分子动力学模拟的结果证实，MOF 1 通过π-π堆积和氢键以及额外的静电相互作用吸收羧基荧光素（FAM）标记的和 5（6）-羧基罗丹明，三乙基铵盐（ROX）标记的探针 ss-DNA（探针 DNA，P-DNA），形成具有淬灭 FAM 和 ROX 荧光的 P-DNA@1 传感平台。在存在靶向埃博拉病毒保守 RNA 序列或埃博拉病毒编码的 miRNA 样片段的情况下，荧光标记的 P-DNA 与分析物杂交形成 P-DNA@RNA 双链体，并从 MOF 1 中释放出来，引发荧光恢复。通过单同步荧光分析也实现了两种靶 RNA 的同时检测。形成的传感平台对埃博拉病毒保守 RNA 序列和埃博拉病毒编码的 miRNA 样片段具有高灵敏度，检测限为皮摩尔级，并且两种探针之间没有交叉反应，具有高选择性。因此，MOF 1 有望成为同时检测两种埃博拉病毒相关序列的有效荧光传感平台，并提高埃博拉病毒病的诊断准确性。
