Sato Shinobu, Takenaka Shigeori
Department of Applied Chemistry, Faculty of Engineering, Kyushu Institute of Technology, Kitakyushu, Fukuoka, Japan.
Anal Sci. 2012;28(7):643-9. doi: 10.2116/analsci.28.643.
An electrochemical DNA detection format has been developed to allow simple and rapid assays with downsized instruments. Since naphthalene diimide derivatives are known to bind to double stranded DNA (dsDNA) with threading intercalation, they are stabilized by pseudo-catenane formation. For example, ferrocenylnaphthalene diimide (FND) can be efficiently concentrated on dsDNA, thus enabling its electrochemical detection. Since dsDNA is formed between single stranded target DNA and DNA probe, the target DNA can be detected electrochemically, using DNA probe-immobilized on the electrode. Further stabilization of naphthalene diimide derivatives as a complex with dsDNA on the electrode is expected to lead to more precise and selective detection of target DNA. This approach was realized by the formation of inclusion complexes of ferrocene with β-cyclodextrin (β-CD) for the FND-dsDNA complexes on the electrode. Combination of adamantylnaphthalene diimide (AND) and ferrocenyl-β-CD (Fc-CD) gave new supramolecular DNA detection assays coupled with DNA probe-immobilized electrodes. The electrochemical signal increased only upon formation of the Fc-CD/AND complex bound to dsDNA on the electrode. Naphthalene diimide carrying ferrocene and β-CD realized "signal on" type detection of dsDNA in homogenous solution. Naphthalene diimide carrying two dithiolene termini provided a new method to immobilize dsDNA on the gold electrode that may be suitable for the electrochemical detection of target DNA.
一种电化学DNA检测形式已被开发出来,以实现使用小型仪器进行简单快速的检测。由于已知萘二酰亚胺衍生物通过穿线插入与双链DNA(dsDNA)结合,它们通过假连环体的形成而得到稳定。例如,二茂铁基萘二酰亚胺(FND)可以有效地富集在dsDNA上,从而实现其电化学检测。由于dsDNA是在单链靶标DNA和DNA探针之间形成的,因此可以使用固定在电极上的DNA探针通过电化学方法检测靶标DNA。预计萘二酰亚胺衍生物与电极上的dsDNA形成的复合物能进一步稳定,从而实现对靶标DNA更精确和选择性的检测。这种方法是通过在电极上为FND-dsDNA复合物形成二茂铁与β-环糊精(β-CD)的包合物来实现的。金刚烷基萘二酰亚胺(AND)和二茂铁基-β-环糊精(Fc-CD)的组合产生了与固定有DNA探针的电极相结合的新型超分子DNA检测方法。只有当电极上与dsDNA结合的Fc-CD/AND复合物形成时,电化学信号才会增加。携带二茂铁和β-CD的萘二酰亚胺实现了在均相溶液中对dsDNA的“信号开启”型检测。携带两个二硫纶末端的萘二酰亚胺提供了一种将dsDNA固定在金电极上的新方法,这可能适用于靶标DNA的电化学检测。