Fang Zhichao, Kelley Shana O
Leslie Dan Faculty of Pharmacy, Department of Pharmaceutical Sciences, and Faculty of Medicine, Department of Biochemistry, University of Toronto, Ontario M5S 3M2, Canada.
Anal Chem. 2009 Jan 15;81(2):612-7. doi: 10.1021/ac801890f.
We report an electrochemical nucleic acids sensing system that exhibits high sensitivity and specificity when challenged with heterogeneous samples of RNA. The platform directly detects specific RNA sequences in cellular and clinical samples without any sample labeling or PCR amplification. The sensor features an electrode platform consisting of three-dimensional gold nanowires, and DNA or RNA hybridization is detected using an electrocatalytic reporter system. In this study, probes made of peptide nucleic acid (PNA) are used to detect a newly identified cancer biomarkera gene fusion recently associated with prostate cancer. The system is able to detect the fusion sequence with 100 fM sensitivity, and retains high sensitivity even in the presence of a large excess of non-complementary sequences. Moreover, the sensor is able to detect the fusion sequence in as little as 10 ng of mRNA isolated from cell lines or 100 ng total RNA from patient tissue samples. The PNA-nanowire nucleic acids sensor described is one of the first electrochemical sensors to directly detect specific mRNAs in unamplified patient samples.
我们报道了一种电化学核酸传感系统，当用RNA的异质样品进行测试时，该系统表现出高灵敏度和特异性。该平台可直接检测细胞和临床样品中的特定RNA序列，无需任何样品标记或PCR扩增。该传感器具有一个由三维金纳米线组成的电极平台，并使用电催化报告系统检测DNA或RNA杂交。在本研究中，由肽核酸（PNA）制成的探针用于检测一种新发现的癌症生物标志物——一种最近与前列腺癌相关的基因融合。该系统能够以100 fM的灵敏度检测融合序列，即使在存在大量非互补序列的情况下也能保持高灵敏度。此外，该传感器能够在从细胞系分离的低至10 ng的mRNA或患者组织样品的100 ng总RNA中检测到融合序列。所描述的PNA-纳米线核酸传感器是首批直接检测未扩增患者样品中特定mRNA的电化学传感器之一。
