Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, QLD, 4072, Australia.
School of Chemistry and Molecular Biosciences, The University of Queensland, QLD, 4072, Australia.
Small. 2016 Dec;12(45):6233-6242. doi: 10.1002/smll.201602161. Epub 2016 Sep 26.
Cancer is a heterogeneous disease which manifests as different molecular subtypes due to the complex nature of tumor initiation, progression, and metastasis. The concept of precision medicine aims to exploit this cancer heterogeneity by incorporating diagnostic technology to characterize each cancer patient's molecular subtype for tailored treatments. To characterize cancer molecular subtypes accurately, a suite of multiplexed bioassays have currently been developed to detect multiple oncogenic biomarkers. Despite the reliability of current multiplexed detection techniques, novel strategies are still needed to resolve limitations such as long assay time, complex protocols, and difficulty in interpreting broad overlapping spectral peaks of conventional fluorescence readouts. Herein a rapid (80 min) multiplexed platform strategy for subtyping prostate cancer tumor and urine samples based on their RNA biomarker profiles is presented. This is achieved by combining rapid multiplexed isothermal reverse transcription-recombinase polymerase amplification (RT-RPA) of target RNA biomarkers with surface-enhanced Raman spectroscopy (SERS) nanotags for "one-pot" readout. This is the first translational application of a RT-RPA/SERS-based platform for multiplexed cancer biomarker detection to address a clinical need. With excellent sensitivity of 200 zmol (100 copies) and specificity, we believed that this platform methodology could be a useful tool for rapid multiplexed subtyping of cancers.
癌症是一种异质性疾病，由于肿瘤起始、进展和转移的复杂性质，表现为不同的分子亚型。精准医学的概念旨在利用这种癌症异质性，通过纳入诊断技术来描述每个癌症患者的分子亚型，从而进行针对性治疗。为了准确表征癌症的分子亚型，目前已经开发了一系列多重生物测定法来检测多种致癌生物标志物。尽管当前的多重检测技术具有可靠性，但仍需要新的策略来解决一些局限性，如检测时间长、复杂的方案以及难以解释传统荧光读数中广泛重叠的光谱峰。本文提出了一种基于 RNA 生物标志物谱的快速（80 分钟）前列腺癌肿瘤和尿液样本分型的多重平台策略。该策略通过将目标 RNA 生物标志物的快速多重等温逆转录-重组酶聚合酶扩增（RT-RPA）与表面增强拉曼光谱（SERS）纳米标签相结合，实现了“一键式”读出。这是 RT-RPA/SERS 平台在用于满足临床需求的多重癌症生物标志物检测中的首次转化应用。该平台具有优异的灵敏度（200 zmol（100 拷贝））和特异性，我们相信该平台方法可能是快速多重癌症分型的有用工具。
