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

描述（由申请人提供）：细菌选择剂的故意释放的威胁促使人们需要快速、无培养的诊断，这种诊断可以在现场或仅具有基本诊断能力的实验室中进行。光纤生物传感器是一种有吸引力的设备，可以检测、记录和传输有关生理变化或环境中各种生物和化学材料存在的信息。这些设备可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、便携式的检测设备，该设备将作为一种平台技术，用于一系列医学相关和临床相关的疾病靶点，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类选择性病原体土拉弗朗西斯菌、鼻疽伯克霍尔德菌和B类。假鼻疽，作为该技术可适用于多种其他试剂和生物标记物的原理的证明。我们的假设是，这些细菌的DNA的特定抗原表位或区域可以在现场或在基本的实验室环境中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偶联到DNA探针以检测特定寡核苷酸序列的DNA杂交测定法;（ii）开发一种使用纳米级光纤生物传感器偶联到抗体以检测特定抗原的抗原结合测定法;和（iii）提高灵敏度并优化纳米级光纤生物传感器的特异性，细菌此外，我们将使用实时PCR和乳胶凝集试验作为对照试验，以比较基于DNA或基于抗原的光纤生物传感器试验的灵敏度和特异性。与表面等离子体共振等竞争技术相比，这种新的生物工程光纤检测方法坚固耐用，便携，价格低廉。这些传感器将填补传染性病原体快速、无培养诊断的重要空白，特别是在主要需要简单便携设备的地区。公共卫生相关性：我们建议开发灵敏、特异、快速的纳米光纤生物传感器，用于诊断A类和B类选择性病原体土拉弗朗西斯菌、鼻疽伯克霍尔德菌和B类。假鼻疽与表面等离子体共振等竞争技术相比，这种新的生物工程光纤检测方法坚固耐用，便携，价格低廉。这些传感器将填补A类和B类选择剂的快速诊断中存在的重要空白，但可以广泛应用。因此，这些测定将作为其应用于诊断各种生物制剂的原理证明。

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.