Nanoscale Optical Fiber Biosensor Assays for Detection of Category A and B Select

描述(由申请人提供)：故意释放细菌选择试剂的威胁促使他们需要快速、无培养的诊断，这种诊断可以在现场或在只有基本诊断能力的实验室进行。光纤生物传感器是一种很有吸引力的设备，它可以检测、记录和传输关于生理变化或环境中各种生物和化学物质的存在的信息。这些设备可以由非专业人员轻松操作，不需要高真空、高温、有机溶剂或洁净室设施。我们的总体目标是开发一种高灵敏度、便携的检测设备，作为一种平台技术，用于一系列具有医学意义和临床意义的疾病靶标，包括传染病病原体。在这个项目中，我们将开发灵敏、特异和快速的纳米级光纤生物传感器，用于诊断A类和B类选定的图拉氏菌、马来伯克霍尔德氏菌和假鼻疽杆菌，以证明这项技术可以适用于各种其他试剂和生物标记物。我们的假设是，这些细菌的特定抗原表位或DNA区域可以在现场或在基本的实验室环境下使用这些光学生物传感器进行检测，从而能够诊断这些或其他细菌。为了实现我们的目标，我们的具体目标是：(I)开发一种使用纳米级光纤生物传感器与DNA探针耦合来检测特定寡核苷酸序列的DNA杂交分析方法；(Ii)使用纳米级光纤生物传感器与抗体耦合来检测特定抗原的抗原结合分析；以及(Iii)提高纳米级光纤生物传感器检测感染细菌特异性寡核苷酸序列和抗原的灵敏度和特异度。此外，我们将使用实时荧光聚合酶链式反应和乳胶凝集试验作为对照分析，以比较基于DNA和基于抗原的光纤生物传感器检测的敏感性和特异性。与表面等离子体共振等竞争技术相比，这种新型的生物工程光纤检测方法坚固耐用，携带方便，价格低廉。这些传感器将填补在快速、无培养诊断感染性病原体方面存在的一个重要空白，特别是在最需要简单和便携式设备的地区。公共卫生相关性：我们建议开发灵敏、特异和快速的纳米级光纤生物传感器，用于诊断A类和B类精选制剂图拉氏方济氏菌、马来伯克霍尔德氏菌和假鼻疽杆菌。与表面等离子体共振等竞争技术相比，这种新型的生物工程光纤检测方法坚固耐用，携带方便，价格低廉。这些传感器将填补在A类和B类精选试剂快速诊断方面存在的一个重要空白，但可以广泛适用。因此，这些检测方法将作为它们应用于各种生物制剂诊断的原则证明。

DESCRIPTION (provided by applicant): The threat of an intentional release of bacterial select agents prompts the need for their rapid, culture-free diagnosis that can be conducted in the field or in laboratories with only basic diagnostic capabilities. Optical fiber biosensors are attractive devices that can detect, record, and transmit information regarding a physiological change or the presence of various biological and chemical materials in the environment. These devices can be easily operated by non-specialist personnel and do not require high vacuum, high temperature, organic solvents, or clean-room facilities. Our overall goal is to develop a highly sensitive, portable, detection device that will serve as a platform technology for a menu of medically relevant and clinically relevant disease targets, including infectious disease agents. For this project, we will develop sensitive, specific, and rapid nanoscale optical fiber biosensors for the diagnosis of the Category A and B select agents Francisella tularensis, Burkholderia mallei, and B. pseudomallei, as proof of principal that this technology can be applicable to a wide variety of other agents and biological markers. Our hypothesis is that specific antigenic epitopes or regions of the DNA of these bacteria can be detected under field or in a basic laboratory setting with these optical biosensors to enable the diagnosis of these or other bacteria. To accomplish our goal our specific aims are to: (i) develop a DNA-hybridization assay using nanoscale optical fiber biosensors coupled to DNA probes to detect specific oligonucleotide sequences; (ii) develop an antigen-binding assay using nanoscale optical fiber biosensors coupled to antibodies to detect specific antigens; and (iii) enhance the sensitivity and optimize the specificity of the nanoscale optical fiber biosensors for detecting oligonucleotide sequences and antigens specific to infectious bacteria. In addition, we will use Real-Time PCR and latex agglutination assays as control assays to compare the sensitivity and specificity of DNA-based or antigen- based optical fiber biosensor assays. The novel bioengineered optical fiber assay is rugged, portable, and inexpensive when compared to competing technologies such as surface plasmon resonance. These sensors will fill an important void that exists in the rapid, culture-free diagnosis of infectious agents, particularly in regions where simple and portable devices are mostly needed. PUBLIC HEALTH RELEVANCE: We propose to develop sensitive, specific, and rapid nanoscale optical fiber biosensor for the diagnosis of the Category A and B select agents Francisella tularensis, Burkholderia mallei, and B. pseudomallei. The novel bioengineered optical fiber assay is rugged, portable, and inexpensive when compared to competing technologies such as surface plasmon resonance. These sensors will fill an important void that exists in the rapid diagnosis of Category A and B select agents, but can be broadly applicable. Therefore, these assays will act as proof-of principle for their application to the diagnosis of a variety of biological agents.