Institute of Materials Research and Engineering, Agency for Science, Technology and Research (A*STAR), 3 Research Link, Singapore 117602, Singapore.
Biointerphases. 2010 Sep;5(3):FA98-104. doi: 10.1116/1.3496962.
In this study, the authors report that sodium citrate can aggregate hexadecyl-trimethyl-ammonium ion(+)-coated gold nanorods (AuNRs), and nucleic acids of different charge and structure properties, i.e., single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), single-stranded peptide nucleic acid (PNA), and PNA-DNA complex, can bind to the AuNRs and therefore retard the sodium citrate-induced aggregation to different extents. The discovery that hybridized dsDNA (and the PNA-DNA complex) has a more pronounced protection effect than ssDNA (and PNA) allows the authors to develop a homogeneous phase AuNRs-based UV-visible (UV-vis) spectral assay for detecting specific sequences of oligonucleotides (20 mer) with a single-base-mismatch selectivity and a limit of detection of 5 nM. This assay involves no tedious bioconjugation and on-particle hybridization. The simple "set and test" format allows for a highly efficient hybridization in a homogeneous phase and a rapid display of the results in less than a minute. By measuring the degree of reduction in AuNR aggregation in the presence of different nucleic acid samples, one can assess how different nucleic acids interact with the AuNRs to complement the knowledge of spherical gold nanoparticles. Besides UV-vis characterization, transmission electron microscopy and zeta potential measurements were conduced to provide visual evidence of the particle aggregation and to support the discussion of the assay principle.
在这项研究中,作者报告称,柠檬酸钠可以聚集十六烷基三甲基铵离子(+)包覆的金纳米棒(AuNRs),以及具有不同电荷和结构特性的核酸,即单链 DNA(ssDNA)、双链 DNA(dsDNA)、单链肽核酸(PNA)和 PNA-DNA 复合物,可与 AuNRs 结合,因此在不同程度上延迟柠檬酸钠诱导的聚集。杂交双链 DNA(和 PNA-DNA 复合物)比单链 DNA(和 PNA)具有更显著的保护作用的发现,使作者能够开发一种基于 AuNRs 的均相相紫外可见(UV-vis)光谱测定法,用于检测具有单碱基错配选择性和检测限为 5 nM 的特定寡核苷酸(20 个碱基)序列。该测定法不需要繁琐的生物缀合和颗粒内杂交。简单的“设置和测试”格式允许在均相相中进行高效杂交,并在不到一分钟的时间内快速显示结果。通过测量不同核酸样品存在下 AuNR 聚集程度的降低,可以评估不同核酸与 AuNRs 的相互作用方式,从而补充对球形金纳米颗粒的了解。除了 UV-vis 表征外,还进行了透射电子显微镜和zeta 电位测量,以提供粒子聚集的直观证据,并支持对测定原理的讨论。