Discriminating viral and bacterial meningitis infections with iDDS probes

使用 iDDS 探针区分病毒性和细菌性脑膜炎感染

• 批准号: 9253915
• 负责人: DAVID A SHAFER
• 金额: $ 24.26万
• 依托单位: GENETAG TECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-03 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9253915
• 关键词: Antibiotics; Bacterial Meningitis; Biological Assay; Brain; Cerebrospinal Fluid; Characteristics; Clinical; Color; Custom; DNA; Data; Data Quality; Detection; Diagnosis; Diagnostic; Disease; Doctor of Philosophy; Enterococcus; Enterovirus; Etiology; FDA approved; Fluorescence; Freeze Drying; Generations; Genome; Goals; Government Agencies; Gram-Positive Bacteria; Haemophilus influenzae; Healthcare Systems; Hospitals; Hybrids; Individual; Infection; Inflammation; Joint Ventures; Label; Laboratories; Legal patent; Length of Stay; Manufacturer Name; Marketing; Membrane; Meningitis; Methods; Molecular; Neisseria meningitidis; Nucleic Acid Amplification Tests; Outcome; Parasites; Parechovirus; Patient Care; Patients; Performance; Pharmaceutical Preparations; Phase; Physicians; Preparation; Price; Property; Public Health; Reaction; Reagent; Regimen; Resistance; Risk; Sampling; Signal Transduction; Site; Specificity; Specimen; Spinal Cord; Streptococcus pneumoniae; Symptoms; System; Technology; Test Result; Testing; Time; Trichomonas vaginalis; University Hospitals; Vancomycin; Viral; Viral Genome; Viral meningitis; Virus; Virus Diseases; Water; Work; base; cost; cost effective; diagnostic assay; fluorophore; fungus; instrument; instrumentation; interest; microorganism; molecular diagnostics; mortality; multiplex detection; phase 1 study; phase 2 study; prevent; success; validation studies

Discriminating viral and bacterial meningitis infections with iDDS probes Confidential PI: Shafer, David A., PhD PROJECT SUMMARY Rapid and accurate molecular diagnostics assays empower physicians to make informed treatment decisions, especially when a disease state may result from variable causes that are indeterminable without such testing. Meningitis may be caused by infection with microorganisms such as viruses, gram-negative (GN) and gram- positive (GP) bacteria, fungi, or parasites. Most frequently, meningitis results from infection with non-polio enteroviruses (EV) and parechoviruses (PV), which typically self-resolve within 10 days. In contrast, bacterial meningitis has a high mortality rate that approaches 100% if not treated. Patients present with similar symptoms regardless of origin; thus, timely diagnosis of the causative agent is paramount for patient care. Patients are commonly treated with an `empiric' regimen of antibiotics until a definitive diagnosis can be made. Nucleic acid amplification tests (NAATs) reduce diagnosis turnaround times by up to several days compared to standard culturing methods, allowing shortened hospital stays, appropriate use of antibiotics, and a reduced financial burden to patients and the health care system. Currently, there are only two FDA-approved NAATs for diagnosing viral and/or bacterial meningitis. Unfortunately, these tests are only approved for use on the manufacturer's fully automated systems, the cost of which is often prohibitive. Further, these assays detect the highly variable viral genomes at a single target site. Collectively, these limitations highlight the need for an instrument-independent assay that can discriminate between viral and bacterial meningitis, with error-checking properties and confirmatory detection at redundant sites in the viral genomes. GeneTAG Technology, Inc. has developed error-checking DNA Detection Switch (iDDS) probes, which employ a fluorescent labeled probe and a slightly mismatched quencher-labeled antiprobe. In the absence of the intended target, the antiprobes hybridize to the probes, quenching their fluorescence and preventing off- target detection. Recently, we developed assays for detecting EV and PV employing two iDDS probes for each target, providing simultaneous 2-color signaling and automatic confirmation of positive or negative test results. We previously developed iDDS probes for GN and GP detection. Ultimately, we are interested developing an FDA-approved iDDS probe-based meningitis assay in a Sample-ReadyTM, lyophilized plate format that will parallel similar tests developed by our collaborator, BioGX Inc., where all reagents, primers, and probes are included, and the test would only require adding sample and molecular grade water. Aim 1 studies will focus on optimizing the dual-iDDS probe EV and PV assays to perform well in multiplex format with commercial EV and PV strains and patient cerebrospinal fluids, converting the multiplex test to Sample-ReadyTM format, and re- testing the Sample-ReadyTM assay with the original samples. A similar strategy will be employed in Aim 2 to prepare a Sample-ReadyTM GN/GP bacteria assay. Delivering a high fidelity, cost-effective meningitis assay should positively impact meningitis diagnostics and provide a direct benefit to public health.

用IDDS区分病毒和细菌性脑膜炎感染探针机密PI：Shafer，David A.，PhD 项目摘要 快速，准确的分子诊断测定法授权医生做出明智的治疗决定， 尤其是当疾病状态可能是由于不确定的原因而没有进行这种测试而导致的。 脑膜炎可能是由感染的微生物，例如病毒，革兰氏阴性（GN）和革兰氏阴性 阳性（GP）细菌，真菌或寄生虫。最常见的是，脑膜炎导致非polio感染 肠病毒（EV）和聚焦病毒（PV），通常在10天内自我弥补。相反，细菌 脑膜炎的死亡率很高，如果不治疗，则接近100％。患者出现相似 症状无论起源如何；因此，及时诊断病因对于患者护理至关重要。 通常用“经验性”抗生素治疗患者，直到可以做出明确的诊断为止。 与核酸扩增测试（NAATS）相比，诊断周转时间最多减少了几天 标准培养方法，允许缩短住院，适当使用抗生素，并减少 患者和医疗保健系统的财务负担。目前，只有两个FDA批准的NAAT 诊断病毒和/或细菌性脑膜炎。不幸的是，这些测试仅被批准用于 制造商的全自动系统，其成本通常是过高的。此外，这些测定法检测 在单个靶位点处高度可变的病毒基因组。总的来说，这些限制突出了 可以区分病毒和细菌性脑膜炎的仪器独立的测定法，并检查错误检查 病毒基因组中冗余位点的性质和验证性检测。 Genetag Technology，Inc。已开发了检查错误的DNA检测开关（IDDS）探针， 采用荧光标记的探针和略微不匹配的淬灭剂抗果杆。在没有 预期的靶标，抗植物与探针杂交，淬灭其荧光并防止异常 目标检测。最近，我们开发了用于检测EV和PV的测定法，每种IDD 目标，同时提供2色信号传导，并自动确认阳性或阴性测试结果。 我们以前开发了用于GN和GP检测的IDDS探针。最终，我们很感兴趣 以FDA批准的IDDS探针基于样本的溶性板板格式的基于探针的脑膜炎测定 我们的合作者Biogx Inc.开发了类似的类似测试，所有试剂，底漆和探针均为 包括，测试只需要添加样品和分子级水。 AIM 1研究将重点 优化双IDDS探针EV和PV测定，以用商业EV和 PV菌株和患者脑脊液的液体，将多重测试转换为样品就绪的格式，并重新 用原始样品测试样品ReadyTM测定法。 AIM 2将采用类似的策略 准备样品ReadyTM GN/GP细菌测定法。提供高保真，具有成本效益的脑膜炎测定 应该对脑膜炎的诊断产生积极影响，并为公共卫生提供直接利益。