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Fluorometric determination of nucleic acids based on the use of polydopamine nanotubes and target-induced strand displacement amplification
Microchimica Acta ( IF 5.7 ) Pub Date : 2018-01-10 , DOI: 10.1007/s00604-017-2632-2
Jia Ge , Dong-Mei Bai , Xin -Geng , Ya-Lei Hu , Qi-Yong Cai , Ke Xing , Lin Zhang , Zhao-Hui Li

AbstractThe authors describe a fluorometric method for the quantitation of nucleic acids by combining (a) cycled strand displacement amplification, (b) the unique features of the DNA probe SYBR Green, and (c) polydopamine nanotubes. SYBR Green undergoes strong fluorescence enhancement upon intercalation into double-stranded DNA (dsDNA). The polydopamine nanotubes selectively adsorb single-stranded DNA (ssDNA) and molecular beacons. In the absence of target DNA, the molecular beacon, primer and SYBR Green are adsorbed on the surface of polydopamine nanotubes. This results in quenching of the fluorescence of SYBR Green, typically measured at excitation/emission wavelengths of 488/518 nm. Upon addition of analyte (target DNA) and polymerase, the stem of the molecular beacon is opened so that it can bind to the primer. This triggers target strand displacement polymerization, during which dsDNA is synthesized. The hybridized target is then displaced due to the strand displacement activity of the polymerase. The displaced target hybridizes with another molecular beacon. This triggers the next round of polymerization. Consequently, a large amount of dsDNA is formed which is detected by addition of SYBR Green. Thus, sensitive and selective fluorometric detection is realized. The fluorescent sensing strategy shows very good analytical performances towards DNA detection, such as a wide linear range from 0.05 to 25 nM with a low limit of detection of 20 pM. Graphical abstractSchematic of a fluorometric strategy for highly sensitive and selective determination of nucleic acids by combining strand displacement amplification and the unique features of SYBR Green I (SG) and polydopamine nanotubes.

中文翻译:

基于使用聚多巴胺纳米管和目标诱导链置换扩增的核酸荧光测定

摘要作者描述了一种通过结合 (a) 循环链置换扩增、(b) DNA 探针 SYBR Green 的独特特征和 (c) 聚多巴胺纳米管来定量核酸的荧光方法。SYBR Green 在插入双链 DNA (dsDNA) 后会发生强烈的荧光增强。聚多巴胺纳米管选择性地吸附单链 DNA (ssDNA) 和分子信标。在没有目标 DNA 的情况下,分子信标、引物和 SYBR Green 被吸附在聚多巴胺纳米管的表面。这导致 SYBR Green 的荧光猝灭,通常在 488/518 nm 的激发/发射波长下测量。在加入分析物(目标 DNA)和聚合酶后,分子信标的茎被打开,以便它可以与引物结合。这会触发目标链置换聚合,在此期间合成 dsDNA。由于聚合酶的链置换活性,杂交的靶标随后被置换。被取代的目标与另一个分子信标杂交。这会触发下一轮聚合。因此,形成了大量 dsDNA,通过添加 SYBR Green 可以检测到。因此,实现了灵敏且选择性的荧光检测。荧光传感策略对 DNA 检测显示出非常好的分析性能,例如从 0.05 到 25 nM 的宽线性范围,检测下限为 20 pM。
更新日期:2018-01-10
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