Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana-Champaign, Illinois 61801, United States.
Anal Chem. 2011 Aug 1;83(15):5949-56. doi: 10.1021/ac201340s. Epub 2011 Jun 29.
In this paper, we present a method for the sensitive detection of microRNAs (miRNAs) utilizing an antibody that specifically recognizes DNA:RNA heteroduplexes and a silicon photonic microring resonator array transduction platform. Microring resonator arrays are covalently functionalized with DNA capture probes that are complementary to solution phase miRNA targets. Following hybridization on the sensor, the anti-DNA:RNA antibody is introduced and binds selectively to the heteroduplexes, giving a larger signal than the original miRNA hybridization due to the increased mass of the antibody, as compared to the 22-mer oligoribonucleotide. Furthermore, the secondary recognition step is performed in neat buffer solution and at relatively higher antibody concentrations, facilitating the detection of miRNAs of interest. The intrinsic sensitivity of the microring resonator platform coupled with the amplification provided by the anti-DNA:RNA antibodies allows for the detection of microRNAs at concentrations as low as 10 pM (350 amol). The simplicity and sequence generality of this amplification method position it as a promising tool for high-throughput, multiplexed miRNA analysis as well as a range of other RNA based detection applications.
在本文中，我们提出了一种利用特异性识别 DNA:RNA 杂合体的抗体和硅光子微环谐振器阵列传输平台来灵敏检测 microRNAs（miRNAs）的方法。微环谐振器阵列通过共价键合与溶液相 miRNA 靶标互补的 DNA 捕获探针进行功能化。在传感器上杂交后，引入抗 DNA:RNA 抗体并选择性地与杂合体结合，由于抗体的质量增加，与 22 个核苷酸的寡核苷酸相比，信号更大。此外，二级识别步骤在纯缓冲溶液中并在相对较高的抗体浓度下进行，有利于检测感兴趣的 miRNAs。微环谐振器平台的固有灵敏度以及抗 DNA:RNA 抗体提供的放大作用，使得能够检测低至 10 pM（350 amol）浓度的 microRNAs。这种放大方法的简单性和序列通用性使其成为高通量、多重 miRNA 分析以及一系列其他基于 RNA 的检测应用的有前途的工具。
