Hierarchical Nanostructuring Array Enhances Mid-Hybridization for Accurate Herbal Identification via ITS2 DNA Barcode
Hierarchical Nanostructuring Array Enhances Mid-Hybridization for Accurate Herbal Identification via ITS2 DNA Barcode
复制标题
分层纳米结构阵列增强中期杂交，通过 ITS2 DNA 条形码进行准确的草药鉴定
DOI：
10.1021/acs.analchem.9b04687
复制
发表时间：
2020-01-21
影响因子：
7.4
通讯作者：
Yang, Fan
中科院分区：
文献类型：
--
作者：
Shi, Ruixue;Hu, Zhigang;Yang, Fan
准确鉴定相近的草药品种，特别是从掺假中鉴别，仍然是一个技术挑战，因为它们的表型和化学成分高度相同。在这里，我们报告了一种直接的，无测序的，高曲率的基于纳米结构的电化学草药传感器（nanoE-herb传感器），使用ITS 2条形码快速准确地识别草药种类。我们设计了一个纳米粗糙的碳支撑的金纳米结构阵列，无石墨烯，一步电沉积。与二维平面相比，三维分形纳米结构表现出高的偏转角，这大大提高了DNA杂交效率，特别是对于中间互补杂交。更重要的是，这种跨尺度阵列生物界面（包括宏观尺度碳和纳米级金分支）可以克服长基因组序列的缓慢扩散的检测障碍和通过突出的3D功能纳米结构的ITS 2二级结构中的隔离变异的不可接近性。我们的nanoE-herb传感器实现了0.18 fM的检测限为64-mer片段的Spectron ITS 2条形码与midhybridization和显示上级特异性，即使是单碱基错配。该传感器还通过直接检测具有ITS 2序列的未纯化的不对称PCR扩增子（类似于500 bp）来精确地区分六种其他掺杂物，这表明其在具有特定DNA条形码的草药物种和病原菌的现场鉴定中具有巨大的潜力。
It remains a technical challenge to accurately identify close species of herbal medicines, especially from adulterants, because of their highly identical phenotypes and chemical compositions. Here, we report a direct, sequencing-free, high-curvature nanostructuring-based electrochemical herb sensor (nanoE-herb sensor) to identify herbal species quickly and accurately using ITS2 barcodes. We engineer a nano-roughened carbon-supported gold nanostructuring array by photolithograph-free, one-step electrodeposition. The 3D fractal nanostructures exhibit a high deflection angle that largely enhances DNA hybridization efficiency, particularly for the midcomplementary hybridization, as compared to the 2D planar surface. More importantly, such a trans-scale array biointerface (including macroscale carbon and nanoscale gold branches) can overcome the detection barrier of slow diffusion of a long genomic sequence and inaccessibility of the sequestered variations in ITS2 secondary structures through the out-protruded 3D functional nanostructures. Our nanoE-herb sensor achieves a detection limit of 0.18 fM for the 64-mer fragment of saffron ITS2 barcode with midhybridization and shows superior specificity against even single-base mismatch. The sensor also precisely differentiates saffron from six other adulterants by directly detecting unpurified asymmetric PCR amplicons (similar to 500 bp) with ITS2 sequences, suggesting its great potential in the field identification of herbal medicinal species and pathogenic bacteria with specific DNA barcodes.