SERS Based Micro-Sensor Arrays for Quantitative miRNAs Detection

基于 SERS 的微传感器阵列用于定量 miRNA 检测

• 批准号: 1064228
• 负责人: Yiping Zhao
• 金额: $ 41万
• 依托单位: University of Georgia Research Foundation Inc
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1064228&HistoricalAwards=false
• 关键词: SERS; Based; Micro; Sensor; Arrays

1064228ZhaoIntellectual MeritSignificant improvements in cancer survival rates are expected with the development of increasingly sensitive and rapid early detection methods and the ability to detect biomarkers of disease. Mounting evidence has implicated microRNA (miRNA) expression profiles as potential cancer biomarkers because aberrant miRNA expression profiles have been linked to the development of several cancer types. Unfortunately, conventional miRNA detection technologies such as microarray analysis and quantitative Reverse Transcription Polymerase Chain Reaction (RT-PCR), while can serve as adequate research tools, require expensive reagents, instrumentation, and multiple time-consuming steps and therefore are not suited for routine miRNA based diagnostic testing. This project is aimed to develop a new platform technology based on surface-enhanced Raman spectroscopy (SERS) for profiling of miRNA expression. This label-free detection method offers a tremendous advantage over conventional miRNA detection methods by eliminating issues associated with species-specific reagents, the need for amplification of the analyte for detection, decreasing expense and complicated procedures that are associated with labelling steps. Central to this technology is a nanofabrication method, oblique angle deposition, recently developed in our laboratory to reproducibly fabricate uniform silver nanorod arrays in large area that dramatically enhance the Raman signal of the molecular signature of the analyte. Preliminary data have been obtained to establish the sensitivity and reproducibility of silver nanorod array SERS substrates and demonstrate rapid (less than 10 second), multiplexed (five different miRNAs) detection and quantization of synthetic miRNA mixtures adsorbed directly (i.e., no hybridization or capture) onto the SERS substrates. In order to achieve the high specificity, high sensitivity, and high throughput detections, this project will optimize the hybridization process of SERS chip to improve sensor?s specificity and sensitivity, as well as to serve a means of concentrating the miRNAs, to develop microarray SERS chip for high sensitive, high specificity and high throughput miRNA detection and multiplexing, to integrate microfluidic SERS chip with electric field concentration to reduce hybridization time and maximize hybridization efficiency and to improve the quality of the quantitative detection, and to demonstrate that the technology can detect and quantify miRNA isolated from cells.Broader ImpactsThis project represents an innovative and comprehensive approach toward developing a platform enabling technology to allow for rapid and sensitive detection of miRNA and miRNA expression profiles. miRNAs are recognized as potential biomarkers of disease, thus a novel sensor platform which advances the ability to detect miRNAs would have unusually high impact on biomedical research by providing the ability to improve our fundamental understanding of miRNAs in gene regulation and in disease pathogenesis. Moreover, a new detection platform would dramatically improve miRNA detection, risk assessment, and treatment and monitoring of patients. These features would provide a new paradigm in RNA biology and offer a translational approach to personal care medicine. A unique educational aspect of this project is the potential to cross-train graduate and undergraduate students in these different scientific disciplines of nanofabrication, spectroscopy and virology. The development of novel viral biosensors provides a mechanism to bridge basic and biomedical research areas, and serves as a unique training vehicle for chemistry, physics and veterinary scientists with interests in viral diseases. In particular, SERS-based biosensors will be used to train a new generation of veterinarians in the College of Veterinary Medicine at University of Georgia to detect infectious diseases using emerging nanotechnology methods.

1064228赵智贤随着越来越灵敏和快速的早期检测方法以及检测疾病生物标志物的能力的发展，预计癌症存活率将显著提高。越来越多的证据表明microRNA（miRNA）表达谱是潜在的癌症生物标志物，因为异常的miRNA表达谱与几种癌症类型的发展有关。不幸的是，常规的miRNA检测技术，如微阵列分析和定量逆转录聚合酶链反应（RT-PCR），虽然可以作为足够的研究工具，但需要昂贵的试剂、仪器和多个耗时的步骤，因此不适合常规的基于miRNA的诊断测试。本项目旨在开发一种基于表面增强拉曼光谱（Sers）的新平台技术，用于分析miRNA表达。这种无标记检测方法通过消除与物种特异性试剂相关的问题、对扩增分析物以用于检测的需要、降低与标记步骤相关的费用和复杂程序，提供了优于常规miRNA检测方法的巨大优势。这项技术的核心是一种纳米纤维方法，斜角沉积，最近在我们的实验室开发的可重复制造均匀的银纳米棒阵列在大面积，显着增强的拉曼信号的分析物的分子签名。已经获得了初步数据以建立银纳米棒阵列Sers基底的灵敏度和再现性，并证明了直接吸附的合成miRNA混合物（即，没有杂交或捕获）到Sers基底上。为了实现高特异性、高灵敏度、高通量的检测，本课题将优化Sers芯片的杂交过程，提高传感器的灵敏度和灵敏度。的特异性和灵敏度，以及作为一种手段，浓缩的miRNA，开发微阵列Sers芯片的高灵敏度，高特异性和高通量的miRNA检测和多重，集成微流控Sers芯片与电场浓度，以减少杂交时间和最大限度地提高杂交效率，并提高质量的定量检测，更广泛的影响该项目代表了一种创新和全面的方法，旨在开发一种平台，使技术能够快速和灵敏地检测miRNA和miRNA表达谱。miRNA被认为是潜在的疾病生物标志物，因此，一种新的传感器平台，提高了检测miRNA的能力，将通过提供提高我们对miRNA在基因调控和疾病发病机制中的基本理解的能力，对生物医学研究产生异常高的影响。此外，新的检测平台将大大改善miRNA检测、风险评估以及患者的治疗和监测。这些特征将为RNA生物学提供一个新的范例，并为个人护理医学提供一种转化方法。这个项目的一个独特的教育方面是交叉培训研究生和本科生在纳米纤维，光谱学和病毒学这些不同的科学学科的潜力。新型病毒生物传感器的开发提供了一种连接基础和生物医学研究领域的机制，并为对病毒性疾病感兴趣的化学，物理和兽医科学家提供了独特的培训工具。特别是，基于SERS的生物传感器将用于培训格鲁吉亚大学兽医学院的新一代兽医，以使用新兴的纳米技术方法检测传染病。