Universal nucleic acid amplification tests-Diagnostics Hub1

通用核酸扩增试验(NAAT)平台依赖于保守和可变的发现 可以利用所有人类病原体中包含的序列来开发强大的快速诊断 方法：研究方法。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 规划该中心的项目，并建立两用设施，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-16S rRNA聚合酶链式反应(16S)的速度，以及 对IBIS T-5000(T-5000)进行基因分型并鉴定多种病原体。该计划将有6个具体的 目标：具体目标1：优化样品制备方法(全血和腹腔液) 提取核酸并确定16S和T-5000在模型中的分析和临床准确性， 动物样本和储存的人类样本。具体目标2：评估16S和T-T的诊断准确性 MARCE-2合作者感兴趣的生物的5000种方法，包括那些入侵地球的生物 黏膜(方案II)或呼吸道(方案III)并引起败血症或腹膜炎； 使用已建立的和新的后置扩增方法区分菌株和血清型的能力。 具体目标3：开发定制的生物信息学软件面板，并确定其临床实用性， T-5000，用于病原体的快速检测、鉴定和基因分型(常见、生物硫酸盐[BT]和 正在出现)，它们入侵并导致呼吸道疾病。T-5000的运力将为 决心确定混合文化和在基础水平上提供基因鉴定的能力。具体目标4： 将16S和/或T-5000应用于正在开发疫苗的MARCE计划项目，并 在动物模型中进行了测试。由于MARCE-2中的几个项目将包括发现、开发和测试 广谱肠道疫苗病原体(尤其是低致病菌)的测定 在接种疫苗后受到挑战的动物)可能是必要的。在后来的几年里，筛查 在组织培养中为不定因素培养的疫苗将在T-5000之前完成。具体目标5： 开发经良好制造规范(GMP)验证的协议，以生产T-5000诊断试剂盒 可用于医院、国家实验室、疾病控制中心和 预防(疾控中心)和世界卫生组织(世卫组织)参考实验室，以及；努力提交 新的专利和寻求16S相关检测的许可商业机会。制造业和 质量人员将准备开发试剂盒检测试剂，其中包括参与建造 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.