School of Natural Sciences, Griffith University, Nathan Campus, QLD 4111, Australia; Queensland Micro- and Nanotechnology Centre, Griffith University, Nathan Campus, QLD 4111, Australia.
Queensland Micro- and Nanotechnology Centre, Griffith University, Nathan Campus, QLD 4111, Australia; Australian Institute for Innovative Materials (AIIM), University of Wollongong, Squires Way, Innovation Campus, North Wollongong, NSW 2500, Australia.
Biosens Bioelectron. 2018 Mar 15;101:275-281. doi: 10.1016/j.bios.2017.09.027. Epub 2017 Sep 20.
A crucial issue in microRNA (miRNA) detection is the lack of sensitive method capable of detecting the low levels of miRNA in RNA samples. Herein, we present a sensitive and specific method for the electrocatalytic detection of miR-107 using gold-loaded nanoporous superparamagnetic iron oxide nanocubes (Au-NPFeONC). The target miRNA was directly adsorbed onto the gold surfaces of Au-NPFeONC via gold-RNA affinity interaction. The electrocatalytic activity of Au-NPFeONC was then used for the reduction of ruthenium hexaammine(III) chloride (RuHex, [Ru(NH)]) bound with target miRNA. The catalytic signal was further amplified by using the ferri/ferrocyanide [Fe(CN)] system. These multiple signal enhancement steps enable our assay to achieve the detection limit of 100aM which is several orders of magnitudes better than most of the conventional miRNA sensors. The method was also successfully applied to detect miR-107 from cancer cell lines and a panel of tissue samples derived from patients with oesophageal squamous cell carcinoma with excellent reproducibility (% RSD = < 5%, for n = 3) and high specificity. The analytical accuracy of the method was validated with a standard RT-qPCR method. We believe that our method has the high translational potential for screening miRNAs in clinical samples.
在 miRNA(microRNA)检测中,一个关键问题是缺乏能够检测 RNA 样本中低水平 miRNA 的敏感方法。在此,我们提出了一种使用负载金的纳米多孔超顺磁性氧化铁纳米立方(Au-NPFeONC)用于电催化检测 miR-107 的灵敏且特异的方法。靶 miRNA 通过金-RNA 亲和相互作用直接吸附到 Au-NPFeONC 的金表面上。然后,Au-NPFeONC 的电催化活性用于还原与靶 miRNA 结合的钌六氨(III)氯化物(RuHex,[Ru(NH) ])。通过使用铁氰化亚铁/亚铁氰化物 [Fe(CN)] 体系进一步放大催化信号。这些多重信号增强步骤使我们的测定法能够实现 100aM 的检测限,比大多数常规 miRNA 传感器好几个数量级。该方法还成功地用于从癌细胞系和一组来自患有食管鳞状细胞癌的患者的组织样本中检测 miR-107,具有出色的重现性(%RSD < 5%,n = 3)和高特异性。该方法的分析准确性通过标准 RT-qPCR 方法进行了验证。我们相信,我们的方法具有在临床样本中筛选 miRNAs 的高转化潜力。