Development of DNA Logic Operations for Viral Diagnostics

描述（由申请人提供）：抗生素的过度使用导致美国抗生素耐药率飙升。用于上呼吸道感染（例如呼吸道合胞病毒（RSV））的抗生素处方比任何其他适应症都要多。医生自我报告称这些处方不适合病毒诊断。快速诊断测试将为医生提供避免不当使用抗生素所需的信息。由于其独特的特异性和可塑性，基于抗体的策略仍然是许多病毒检测平台的关键组成部分。然而，基于高灵敏度抗体的检测技术的一个主要障碍是抗体和非靶标之间固有地存在非特异性相互作用。该申请描述了一种将 DNA 报告标签与 DNA 抗体标签之间的逻辑运算相结合的方法，以开发基于 DNA 标签的病毒检测的新范例。在所提出的方法中，不是指定唯一的、但任意选择的DNA序列作为DNA抗体标签，而是特意设计DNA序列以促进标签之间的逻辑操作。在目标 1 中，这些“DNA 逻辑标签”将被设计、测试，并与金纳米粒子表面的抗体共同偶联。据推测，这种逻辑标签方法将允许使用逻辑 NOT 操作控制非特异性抗体相互作用（目标 2），以及使用逻辑 OR 和标签之间的操作独立微调检测灵敏度和特异性的方法（目标 3）。这些假设将通过测量逻辑运算对在日益复杂的生物环境中进行的 RSV 检测测定的灵敏度和特异性的影响来评估。高度敏感和特异性的诊断工具（例如这里提出的）将在即时诊断中变得越来越重要，以快速识别病原体并帮助指导选择适当的治疗方法。 公共卫生相关性：用于上呼吸道感染的抗生素处方（估计每年 1 亿张处方）比任何其他适应症都要多，医生自我报告称，如果有病毒诊断，这些处方将是不合适的。迫切需要快速、简单且准确的诊断测试来及早发现呼吸道病毒感染。该项目的目标是评估一种新的病毒检测技术，用于医学上重要的范例呼吸道合胞病毒。

DESCRIPTION (provided by applicant): The overuse of antibiotics has led to soaring rates of antibiotic resistance in the U.S. More antibiotic prescriptions are written for upper respiratory infections, such as respiratory syncytial virus (RSV), than for any other indication. Physicians self-report that these prescriptions would be inappropriate for a viral diagnosis. Rapid diagnostic tests would give physicians the information needed to avoid inappropriate antibiotic use. Because of their unique specificity and plasticity, antibody-based strategies continue to be a key component of many virus detection platforms. However, a major stumbling block of high-sensitivity antibody-based detection technologies is the innate presence of non-specific interactions between antibodies and non-targets. This application describes a method to combine DNA reporter tags and logical operations among DNA-antibody tags to develop a new paradigm in DNA tag-based virus detection. In the proposed approach, instead of specifying unique, but arbitrarily chosen DNA sequences as DNA-antibody tags, the DNA sequences are purposely designed to facilitate logical operations among tags. In Aim 1 these "DNA logic tags" will be designed, tested, and co-coupled with antibodies to gold nanoparticle surfaces. It is hypothesized that this logic tag approach will permit control of non-specific antibody interactions using a logical NOT operation (Aim 2), and a means to independently fine-tune detection sensitivity and specificity using logical OR and operations among tags (Aim 3). These hypotheses will be evaluated by measurements of the effect of the logical operations on the sensitivity and specificity of an RSV detection assay performed in an increasingly complex biological environment. Highly sensitive and specific diagnostic tools, such as proposed here, will become increasingly important in point-of-care diagnostics to rapidly identify pathogens and help guide the choice of appropriate therapeutic treatment. PUBLIC HEALTH RELEVANCE: More antibiotic prescriptions are written for upper respiratory infections (an estimated 100 million prescriptions a year) than for any other indication, and doctors self-report that these prescriptions would be inappropriate if a viral diagnosis was available. Rapid, simple and accurate diagnostic tests for early detection of respiratory virus infections are sorely needed. The goal of this project is to evaluate a new viral detection technology for the medically important exemplar, respiratory syncytial virus.