Rapid, Label-free, Multiplex Detection of MicroRNAs

描述(申请人提供)：全世界迫切需要用于检测和监测恶性肿瘤的微创测试，以减少癌症造成的发病率和死亡率。虽然前景看好，但基于血液的蛋白质组学分析尚未在许多常见的人类癌症中得到临床验证。因此，正在研究新的办法，这些办法可能会补充或改进现有战略。特别是，microRNAs(MiRNAs)作为一系列疾病的潜在预测生物标记物已经得到了相当大的关注。这个第一阶段的研究项目将评估使用一种名为表面等离子体增强公共路径干涉(SPE-CPI)的新型超灵敏检测技术从血液样本中快速、无标记地识别和定量miRNAs的可行性。SPE-CPI与标准的表面等离子体共振(SPR)检测方法(如Biacore)的不同之处在于，它结合了表面等离子体共振的高灵敏度与CPI单光束路径的更高稳定性和更低噪声。SPE-CPI还允许同时监测单个样品中的多种miRNA。在第一阶段，基于SPE-CPI的在有血清存在的情况下测量癌症相关miRNAs的协议将被识别和合格。特异性和检测限将是这项可行性证明研究的关键参数。此外，使用优化的SPE-CPI方法，我们将使用肺癌患者和健康人的血清样本来评估SPE-CPI作为诊断工具检测miRNAs的潜力。如果成功，这项研究将在癌症以及可能由miRNA调节的其他疾病的新诊断策略方面向前迈进一大步。无标记传感技术的进步还将在低丰度蛋白质测量、蛋白质-蛋白质相互作用研究、传染病检测、生物病毒病原体检测、蛋白质-药物相互作用、食品安全、化学污染物检测等方面打开更广泛的应用。将在这些研究中评估的SPE-CPI仪器体积小、价格便宜，并且只需要最少的样本操作即可进行分析。负担得起、易于使用的探测器用于在医疗机构(即医生办公室)进行快速筛查，将是卫生保健行业的重大突破。 公共卫生相关性：这个小型企业创新研究(SBIR)第一阶段项目将评估一种用于血清样本中肺癌生物标记物的非侵入性多重定量的超灵敏检测技术。如果成功，这项研究将是癌症和其他疾病的护理点诊断新战略的重要一步。生物传感器技术的进步还将在低丰度蛋白质测量、蛋白质-蛋白质相互作用研究、传染病检测、生物病毒病原体检测、蛋白质-药物相互作用、疫苗开发、食品安全、化学污染物检测等方面开辟更广泛的应用。

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.