Rapid Disease Diagnostics using Photonic Crystal Enhanced Antigen Biomarker

血液中疾病生物标志物的常规高灵敏度多路检测将对 早期诊断和治疗选择。一种新的高效的广义类别检测方法 对疾病的研究是使用蛋白质微阵列对抗体进行多路检测。作为数量更多的抗体 生物标志物正在被识别和临床验证，用于病毒感染、癌症、心血管疾病和 自身免疫性疾病，迫切需要以更高的灵敏度和更高的信噪比来检测它们 同时，能够以较低的每次测试成本执行自动化生物标记物分析。在 在提出的项目中，研究人员将光子晶体增强荧光(PCEF)技术与 用于多重微斑点荧光夹心检测的新型尺寸排除血液过滤技术 临床实验室环境平台。我们的方法将几项创新集成到 廉价的塑料传感器墨盒和桌面检测仪器：1.硅基 将使用集成了法布里-珀罗光学腔的纳米结构共振光学光子晶体芯片 提供比当前一代PCEF设备高约50倍的信号增强，后者已经成为常规设备 仅用10L样品体积，在复杂介质中提供1 pg/ml的检出限。2.激光加工 血液过滤器将在60秒内从一滴肝素化的全血中分离出血浆，使整个 无需用户干预即可在&lt；60分钟内自动执行分析方案。3.一款创新的激光 扫描方法用于将来自小型半导体激光器的光直接耦合到PC表面 最佳的“开共振”条件，使仪器坚固、紧凑，成本为10K美元。4.A 统计生物信息学工具，指示测试样本中生物标志物的存在或不存在。作为一种 该系统的应用实例，我们将集中精力检测一组3种血清抗体 人乳头瘤病毒(HPV)作为识别口咽部易感性较高患者的手段 癌症(OPC)，尽管同样的技术可以扩展到检测广泛类别的抗体 生物标志物。新仪器将首先在全血中加入生物标记物进行测试，并得出结果 在微孔板和Luminex微珠系统中与单抗ELISA进行比较。进一步 将对已知HPV患者的临床血液样本进行验证和比较 曝光。我们的长期目标是演示一个可以广泛应用于多路传输的原型系统 血清抗体生物标志物分析。

Routine high-sensitivity multiplexed detection of disease biomarkers in blood will have an impact on early diagnosis and therapy selection. A novel and highly effective approach for detection of broad categories of disease is multiplexed detection of antibodies using protein microarrays. As a greater number of antibody biomarkers are being identified and clinically validated for viral infection, cancer, cardiovascular disease, and autoimmune disease, there is a strong need to detect them with greater sensitivity and greater signal-to-noise ratio, while at the same time being able to perform automated biomarker analysis with low cost per test. In the proposed project, the investigators integrate photonic crystal enhanced fluorescence (PCEF) technology with a novel size-exclusion blood filtration technology to develop a multiplexed microspot fluorescent sandwich assay platform for the clinical laboratory environment. Our approach integrates several innovations into an inexpensive plastic-based sensor cartridge and desktop detection instrument: 1. A silicon-based nanostructured resonant optical photonic crystal chip with an integrated Fabry-Perot optical cavity will be used to deliver ~50x greater signal enhancement than current-generation PCEF devices, which already routinely provide <1 pg/ml limits of detection in complex media, using only 10 l sample volumes. 2. A laser-machined blood filter will separate plasma from a droplet of heparinized whole blood in 60 seconds, enabling the entire assay protocol to be performed automatically in <60 minutes without user intervention. 3. An innovative laser scanning approach is used to couple light from a small semiconductor laser directed to the PC surface in the optimal “on-resonance” condition, resulting in a rugged, compact instrument with a cost of <$10K. 4. A statistical bioinformatics tool indicating the presence or absence of the biomarkers in the test sample. As an example application of the system, we will focus our effort on detection of a panel of 3 serum antibodies for human papillomavirus (HPV) as a means for identifying patients with greater susceptibility to oropharyngeal carcinoma (OPC), although the same technology can be extended to detection of broad classes of antibody biomarkers. The new instrument will be first tested upon biomarkers spiked into whole blood, with results compared against single-antibody ELISA in microplates and the Luminex bead-based system. Further validation and comparison will be performed upon clinical blood samples from patients with known HPV exposure. Our long-term goal is to demonstrate a prototype system that can be applied broadly for multiplexed serum antibody biomarker analysis.