Interdepartmental Biological Sciences Program, Northwestern University, 2205 Tech Drive, Evanston, Illinois 60208, USA.
Anal Chem. 2012 May 1;84(9):4153-60. doi: 10.1021/ac3004055. Epub 2012 Apr 10.
We report the development of a novel Scanometric MicroRNA (Scano-miR) platform for the detection of relatively low abundance miRNAs with high specificity and reproducibility. The Scano-miR system was able to detect 1 fM concentrations of miRNA in serum with single nucleotide mismatch specificity. Indeed, it provides increased sensitivity for miRNA targets compared to molecular fluorophore-based detection systems, where 88% of the low abundance miRNA targets could not be detected under identical conditions. The application of the Scano-miR platform to high density array formats demonstrates its utility for high throughput and multiplexed miRNA profiling from various biological samples. To assess the accuracy of the Scano-miR system, we analyzed the miRNA profiles of samples from men with prostate cancer (CaP), the most common noncutaneous malignancy and the second leading cause of cancer death among American men. The platform exhibits 98.8% accuracy when detecting deregulated miRNAs involved in CaP, which demonstrates its potential utility in profiling and identifying clinical and research biomarkers.
我们开发了一种新颖的扫描式 microRNA(Scano-miR)平台,用于高特异性和高重复性地检测相对低丰度的 microRNA。Scano-miR 系统能够在血清中以单核苷酸错配特异性检测到 1 fM 浓度的 microRNA。实际上,与基于分子荧光染料的检测系统相比,它为 microRNA 靶标提供了更高的灵敏度,在相同条件下,88%的低丰度 microRNA 靶标无法被检测到。Scano-miR 平台在高密度阵列格式中的应用证明了其在各种生物样本中高通量和多重 microRNA 分析的用途。为了评估 Scano-miR 系统的准确性,我们分析了来自患有前列腺癌(CaP)的男性样本的 microRNA 图谱,CaP 是最常见的非皮肤恶性肿瘤,也是美国男性癌症死亡的第二大主要原因。该平台在检测与 CaP 相关的失调 microRNA 时表现出 98.8%的准确率,这表明其在分析和鉴定临床及研究生物标志物方面具有潜在的应用价值。