Development of Fiber Optic Biosensors for Detection of Cancer Molecular Markers

描述（由申请人提供）： 目前低成本的高通量技术用于癌症的检测和诊断、预测治疗效果或评估临床结局的应用还很有限。认识到这一挑战，美国国立卫生研究院/国家癌症研究所的癌症预防研究小额赠款计划（PAR-08-055）寻求资助与“识别、开发和评估与临床前癌症检测相关的生物分析技术、方法和临床技术”有关的“试点和可行性研究”。直接响应于该PAR，在本申请中，我们提出开发一种新的低成本光纤生物传感器工具（浸渍探针）来测量分子生物标志物，其可用于癌症的早期检测/诊断、监测治疗反应或预测临床结果。该装置将依赖于锥形光纤生物传感器（CTFOB）探头和二极管激光激发的组合，并将根据夹心免疫测定原理发挥作用。分析物的量将以荧光信号的形式测量。荧光信号将由倏逝波激发产生。此外，可以使用几个蘸取探针来测量单个样品中的多个标志物。此外，我们的初步研究结果表明，该工具能够检测样品中的少量分析物（灵敏度高10倍），与其他目前可用的检测分析系统（包括Western印迹和基于珠的Luminex免疫测定）相比，使用减少量的试剂（少10-100倍）和时间（不到一小时）。我们建议的研究有三个具体目标。具体目标#1将集中于通过优化探针参数（锥角、锥长和探针半径）以及将现有夹心免疫测定技术应用于光纤探针来增强荧光信号。在具体目标#2中，我们将集中于通过采用催化报告沉积（CARD）方法来提高光纤探针的检测速度和灵敏度。该技术将用于检测和定量单个样本（例如血清样本）中的多种标志物。具体目标#3将是优化成本，周转时间和试剂使用的这种技术，并比较它与其他检测技术。这些研究的成功完成导致低成本，便携式光纤工具的开发，这将被用于早期检测，诊断，治疗效果，预后和几种人类恶性肿瘤的监测。

DESCRIPTION (provided by applicant): Currently the low-cost high-throughput technologies for the detection and diagnosis of cancer, predict the efficacy of therapy or assess the clinical outcomes are limited. In recognition of this challenge, the cancer Prevention Research Small Grant Program (PAR-08-055) of the NIH/NCI seeks to fund "pilot and feasibility studies" related to "identification, development, and evaluation of biological analytic techniques, methodologies, and clinical technologies relevant to pre-clinical cancer detection. In direct response to this PAR, in this application we propose to develop a novel low cost fiber optic biosensor tool (dip-probe) to measure the molecular biomarkers which can be utilized for early detection/diagnosis of cancer, monitor the therapy response or predict the clinical outcomes. This device will rely on combination tapered fiber optic biosensor (CTFOB) probe and diode laser excitation, and will function on a sandwich immunoassay principle. The amount of the analyte will be measured in the form of a florescence signal. The fluorescence signal will be generated by evanescent wave excitation. Additionally, several dip-probes can be used to measure multiple markers in a single sample. Furthermore, our preliminary findings suggest that this tool is capable of detecting small amounts of analytes in samples (10 times higher sensitivity) utilizing reduced amounts of reagents (10-100 folds less) and time (less than an hour) when compared to other currently available detection assay systems including Western blot, and bead-based Luminex immunoassay. Our proposed studies will have three specific aims. Specific aim # 1 will focus on enhancement of fluorescence signal by optimization of probe parameters (tapered angle, taper length and probe radius), and adoption of existing sandwich immunoassay techniques to fiber optic probes. In specific aim # 2 we will focus on increasing the detection speed and sensitivity by adopting catalyzed reporter deposition (CARD) method to fiber optic probes. The technique will be applied to detect and quantify multiple markers in a single sample (e.g. serum sample). Specific aim # 3 will be to optimize the cost, turnaround time and reagent usage of this technique and compare it with other detections techniques. Successful completion of these studies result in the development of a low cost, portable fiber optic tool which will be utilized in early detection, diagnosis, therapy efficacy, prognosis, and surveillance of several human malignancies.