Rapid, Label-free, Multiplex Detection of MicroRNAs

描述（由申请人提供）：世界范围内迫切需要用于检测和监测恶性肿瘤的微创测试，以减少癌症引起的发病率和死亡率。虽然有希望，但基于血液的蛋白质组学分析尚未在临床上验证许多常见的人类癌症。因此，正在研究可能补充或改进目前战略的新办法。特别是，microRNAs （miRNAs）作为许多疾病的潜在预测生物标志物已经获得了相当大的关注。这个第一阶段的研究项目将评估使用一种被称为表面等离子体增强共程干涉测量（SPE-CPI）的新型超灵敏检测技术对血液样本中的mirna进行快速、无标记鉴定和定量的可行性。SPE-CPI不同于标准的表面等离子体共振（SPR）检测方法，如Biacore，它结合了SPR的高灵敏度和CPI单光束路径的高稳定性和低噪声。SPE- CPI还允许在单个样品中同时监测多种miRNA。在I期，基于SPE-CPI的检测血清中癌症相关mirna的方案将被确定和鉴定。特异性和检测限将是这个证明可行性研究的关键参数。此外，使用优化的SPE-CPI方法，我们将评估SPE-CPI作为检测肺癌患者与健康个体血清样本中mirna的诊断工具的潜力。如果成功，这项研究将是癌症和其他可能由miRNA调节的疾病的新诊断策略向前迈出的重要一步。无标签传感技术的进步也将为低丰度蛋白质的测量、蛋白质-蛋白质相互作用研究、传染病检测、生物战病原体检测、蛋白质-药物相互作用、食品安全、化学污染物检测等开辟更广泛的应用。将在这些研究中评估的SPE-CPI仪器体积小，价格低廉，并且需要最少的样本操作进行分析。负担得起、易于使用的检测器，可在护理设施现场（即医生办公室）进行快速筛查，这将是医疗保健行业的一项重大突破。

DESCRIPTION (provided by applicant): Minimally invasive tests for the detection and monitoring of malignancies are urgently needed worldwide to reduce the morbidity and mortality caused by cancer. While promising, blood-based proteonomic assays have yet to be clinically validated for many common human cancers. Accordingly, new approaches are being investigated that will likely complement or improve on current strategies. In particular, microRNAs (miRNAs) have gained considerable attention as potentially predictive biomarkers for a number of disorders. This Phase I research project will evaluate the feasibility of rapid, label-free identification and quantification of miRNAs from blood samples using a novel ultra-sensitive detection technology known as Surface Plasmon Enhanced Common Path Interferometry (SPE-CPI). SPE-CPI differs from standard Surface Plasmon Resonance (SPR) detection methods, such as Biacore, in that it combines the high sensitivity of SPR with the higher stability and lower noise of a CPI single-beam path. SPE- CPI also allows for the simultaneous monitoring of multiple species of miRNA in a single sample. During Phase I, SPE-CPI based protocols for measuring cancer-related miRNAs in the presence of serum will be identified and qualified. Specificity and limit of detection will be critical parameters for this proof-of-feasibility study. Further, using the optimized SPE-CPI methods, we will evaluate the potential of SPE-CPI as a diagnostic tool for the detection of miRNAs using serum samples from patients with lung cancer versus healthy individuals. If successful, this research will be a major step forward in new diagnostic strategies for cancer and possibly other diseases that may be modulated by miRNA. Advancements in label-free sensing technologies will also open broader applications for the measurement of low abundance proteins, protein- protein interaction studies, infectious disease detection, biowarfare pathogen detection, protein- drug interactions, food safety, detection of chemical pollutants, and the like. The SPE-CPI instrument that will be evaluated in these studies is small, inexpensive, and requires minimal sample manipulation for analysis. Affordable, easy-to use detectors for rapid screening at point of care facilities, i.e. doctors' offices, would be a significant breakthrough in the health care industry. PUBLIC HEALTH RELEVANCE: This Small Business Innovation Research (SBIR) Phase I project will evaluate an ultra-sensitive detection technology for the non-invasive multiplex quantification of lung cancer biomarkers in serum samples. If successful, this research will be a major step forward in a new strategy for point of care diagnosis of cancer and other diseases. Advancements in biosensor technologies will also open broader applications for the measurement of low abundance proteins, protein-protein interaction studies, infectious disease detection, biowarfare pathogen detection, protein-drug interactions, vaccine development, food safety, detection of chemical pollutants, and the like.