Visual detection of single-base mismatches in DNA using hairpin oligonucleotide with double-target DNA binding sequences and gold nanoparticles.
Visual detection of single-base mismatches in DNA using hairpin oligonucleotide with double-target DNA binding sequences and gold nanoparticles.
复制标题
DOI：
10.1016/j.bios.2011.12.055
复制
发表时间：
2012-04-15
影响因子：
12.6
通讯作者：
Liu, Guodong
中科院分区：
文献类型：
--
作者：
He, Yuqing;Zhang, Xibao;Zhang, Sanquan;Kris, Mak Ka Long;Man, Fong Chi;Kawde, Abdel-Nasser;Liu, Guodong
我们描述了一种发夹结构的寡核苷酸（HO），其环中具有双靶DNA结合序列，茎中具有11个碱基，用于视觉检测DNA中的单碱基错配（SBM），具有高度特异性。巯基修饰的HO通过自组装过程固定在金纳米粒子（Au-NP）表面。检测SBM的策略依赖于HO对完全匹配的DNA和SBM的独特分子识别特性，以在Au-NP表面产生不同量的双链DNA，通过双链DNA与预固定的DNA探针之间的DNA杂交反应将其捕获在侧向流动测试条的测试区上。Au-NP的积累产生特征性的红色条带，使得能够视觉检测SBM。研究发现，通过在HO环中引入双靶DNA结合序列，HO区分完全匹配DNA和SBM的能力显著提高。完全匹配的DNA与SBM之间的信号比高达28，这远远高于常规的HO或分子信标。应用该方法检测常染色体隐性遗传性先天性鱼鳞病患者转氨酶1基因Arg 142 Cys和Gly 529 Ile突变位点。研究结果表明，这种新的HO是一种潜在的分子识别探针，可用于核酸生物传感器和生物检测。该方法可用于遗传疾病的即时诊断和检测传染性病原体或警告生物战剂。
We describe a hairpin oligonucleotide (HO) with double-target DNA binding sequences in the loop and 11-base in the stem for visual detection of single-base mismatches (SBM) in DNA with highly specificity. The thiol-modified HO was immobilized on gold nanoparticle (Au-NP) surface through a self-assembling process. The strategy of detecting SBM depends on the unique molecular recognition properties of HO to the perfect-matched DNA and SBM to generate different quantities of duplex DNA on the Au-NP surface, which are captured on the test zone of lateral flow test strip via the DNA hybridization reaction between the duplex DNA and preimmobilized DNA probe. Accumulation of Au-NPs produces the characteristic red bands, enabling visual detection of SBM. It was found that the ability of HO to differentiate perfect-matched DNA and SBM was increased dramatically by incorporating double-target DNA binding sequences in the loop of HO. The signal ratio between perfect-matched DNA and SBM was up to 28, which is much higher than that of conventional HO or molecular beacon. The approach was applied to detect the mutation sites, Arg142Cys and Gly529Ile, of transglutaminase 1 gene in autosomal recessive congenital ichthyosis. The results presented here show that the new HO is a potential molecular recognition probe for the future development of nucleic acid-based biosensors and bioassays. The approach can be used for point-of-care diagnosis of genetic diseases and detecting infectious agents or warning against bio-warfare agents.
登录
查看更多内容
影响因子：
46.9
作者：
Dubertret, B;Calame, M;Libchaber, AJ
通讯作者：
Libchaber, AJ
影响因子：
1.8
作者：
Karadeniz, Hakan;Kuralay, Filiz;Erdem, Arzum
通讯作者：
Erdem, Arzum
影响因子：
--
作者：
Kaltenboeck B;Wang C
通讯作者：
Wang C
影响因子：
7.4
作者：
Toubanaki, Dimitra K.;Christopoulos, Theodore K.;Flordellis, Christodoulos S.
通讯作者：
Flordellis, Christodoulos S.
DOI：
10.1007/978-1-60327-411-1_17
发表时间：
2009
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Barreiro LB;Henriques R;Mhlanga MM
通讯作者：
Mhlanga MM