Universal nucleic acid amplification tests-Diagnostics Hub1

通用核酸扩增测试（NAAT）平台依赖于发现保守和可变的核酸扩增测试（NAAT）平台。 包含在所有人类病原体中的序列可用于开发稳健的快速诊断方法， 方法.我们将发展两个已经先进的基础广泛的诊断平台，这两个平台都将支持 在中心规划项目，并建立双重用途的实用程序，即生物防御/新出现的感染和 常见的临床感染，增加了商业化的可能性。互补但 将利用每个平台的独特优势-16 S rRNA PCR（16 S）的速度，以及 IBIS T-5000（T-5000）的基因分型和鉴定广泛的病原体。该计划将有6个具体的 目的：具体目的1：优化用于以下目的的样品制备方法（全血和腹腔液）： 提取核酸并确定16 S和T-5000在模型中的分析和临床准确性， 动物样品和库存的人类样品。具体目标2：评价16 S和T- MARCE-2合作者感兴趣的生物体的5000种方法，包括入侵GI的生物体 粘膜（程序II）或呼吸道（程序III），并导致败血症或腹膜炎;并证明 使用已建立的和新的扩增后方法区分菌株和血清型的能力。 具体目标3：开发生物信息学软件的定制面板，并确定其临床效用， T-5000用于病原体（常见、生物威胁[BT]和 新出现的），其侵入并引起呼吸道疾病。T-5000的能力将是 决心识别混合文化和提供基本水平的基因分型的能力。具体目标4： 将16 S和/或T-5000应用于正在开发疫苗的MARCE计划项目， 在动物模型中测试。由于MARCE-2中的几个项目将包括发现、开发和测试， 对于广谱肠疫苗，确定病原体的存在（特别是那些在低浓度下）， 可能需要在接种疫苗后攻毒的动物中进行检测。在以后的几年里， 在组织培养物中生长的外源因子疫苗将由T-5000完成。具体目标五： 制定良好生产规范（GMP）验证方案，以生产T-5000诊断试剂盒， 商业化，可用于医院，国家实验室，疾病控制中心， 预防中心（CDC）和世界卫生组织（WHO）参考实验室，并努力实现备案 新的专利，并寻求16 S相关分析的许可商业机会。制造和 质量人员将准备试剂盒检测试剂的开发，包括参与构建 并测试试剂的原型运行。

Universal Nucleic Acid Amplification Test (NAAT) platforms rely on the discovery that conserved and variable sequences contained in all human pathogens can be exploited for development of robust rapid diagnostic methods. We will grow two already advanced broad based diagnostic platforms which will both support Program Projects in the Center, and establish utility for dual use, i.e. biodefense/emerging infections and commonly encountered clinical infections, increasing likelihood of commercialization. Complementary but distinct strengths of each platform will be harnessed - speed for the 16S rRNA PCR (16S), and ability to genotype and identify a wide range of pathogens for IBIS T-5000 (T-5000). The program will have 6 specific aims: Specific Aim 1: Optimizing sample preparation methods (whole blood and peritoneal fluid) for extraction of nucleic acids and determine analytic and clinical accuracy of 16S and T-5000 in mock-ups, animal samples and banked human samples. Specific Aim 2: Evaluating diagnostic accuracy of 16S and T- 5000 methods for organisms of interest to MARCE-2 collaborators, including those which invade the Gl mucosa (Program II) or respiratory tract (Program III) and cause sepsis or peritonitis; and demonstrating capacity to differentiate strains and serovars, using both established and novel post-amplification methods. Specific Aim 3: Developing customized panels of bioinformatic software for, and determine clinical utility of, T-5000 for rapid detection, identification, and genotyping of pathogens (common, biothreat [BT] and emerging) which invade and cause disease of the respiratory tract. The capacity of the T-5000 will be determined to identify mixed cultures and the ability to provide genotying at a basic level. Specific Aim 4: Applying the 16S and/or T-5000 to MARCE Programs project for which vaccines are being developed and tested in animal models. Since several projects in MARCE-2 will include discovery, development and testing of broad-spectrum enteric vaccines, determination of the presence of pathogen (especially those at low levels) in animal challenged following vaccine delivery may be necessary. For later years, screening vaccines that are grown in tissue culture for adventitious agents will be done by T-5000. Specific Aim 5: Developing Good Manufacturing Practice (GMP) validated protocols to produce a T-5000 diagnostic kit for commercialization that could be used in hospitals, State laboratories, Centers for Disease Control and Prevention (CDC) and World Health Organization (WHO) reference laboratories, and; to work towards filing new patents and pursuing license commercial opportunities for 16S related assays. Manufacturing and Quality personnel will prepare development of kit testing reagents which will include participation in building and testing prototype runs of reagents.