Impedimetric detection of influenza A (H1N1) DNA sequence using carbon nanotubes platform and gold nanoparticles amplification
Impedimetric detection of influenza A (H1N1) DNA sequence using carbon nanotubes platform and gold nanoparticles amplification
复制标题
DOI：
10.1039/c000532k
复制
发表时间：
2010-01-01
期刊：
影响因子：
4.2
通讯作者：
del Valle, M.
中科院分区：
文献类型：
--
作者：
Bonanni, A.;Pividori, M. I.;del Valle, M.
在这项工作中，我们报告了使用阻抗基因传感器来模型检测 H1N1 猪流感相关 DNA 序列。与目标 H1N1 病毒序列互补的寡核苷酸 DNA 探针通过共价结合固定在电极表面。采用并比较了两种不同的方案，即与 DNA 靶标的直接杂交和夹心方案。在这两种情况下，所得的杂交体都经过生物素标记，以允许与链霉亲和素金纳米颗粒（链霉-AuNP）进行额外的缀合。后者的目的是增强阻抗信号，从而提高该技术的灵敏度。信号放大步骤后使用夹心方案获得的最佳检测限为 7.5 fmol（相当于 577 pmol L-1）。此外，还进行了金增强处理，以比较金纳米粒子在电极表面上的存在和分布。作为另一种可视化方法，采用链霉亲和素共轭量子点 (strept-QD) 来获取 DNA-生物素-链霉-QD 电极表面的荧光图像。最后，从技术的可行性和可行性方面对阻抗和显微镜进行了比较。
In this work we report the use of an impedimetric genosensor for the model detection of H1N1 swine flu correlated DNA sequence. An oligonucleotide DNA probe, complementary to the target H1N1 virus sequence, was immobilized onto the electrode surface by covalent binding. Two different protocols, i.e. direct hybridization with the DNA target and a sandwich scheme, were employed and compared. In both cases the resulting hybrid was biotin-labelled to allow the additional conjugation with streptavidin gold nanoparticles (strept-AuNPs). The latter were used with the aim of enhancing the impedimetric signal, thus improving the sensitivity of the technique. The best limit of detection, obtained with the sandwich scheme after signal amplification step was 7.5 fmol (corresponding to 577 pmol L-1). Furthermore, a gold enhancement treatment was performed in order to compare the presence and distribution of gold nanoparticles onto the electrode surface. As an alternative way of visualization, streptavidin conjugate quantum dots (strept-QD) were employed to obtain fluorescence images of the DNA-biotin-strept-QD electrode surface. Finally, a comparison between impedance and microscopy was performed in terms of viability and feasibility of the techniques.