Rapid Molecular Diagnostic for Chlamydia and Gonorrhea at the Point-of-Care

描述(申请人提供)：美国有400多万人感染了衣原体(CT)和淋病(NG)。这些性传播感染(STI)的早期症状很少，但会导致严重的并发症，从不孕不育到多重抗药性，以及艾滋病毒感染和传播的增加。减少这些性传播感染的传播需要对感染者进行早期诊断和治疗。要取得成功，需要广泛使用低成本、强大和易于使用的分子诊断技术，以便在护理点(POC)最早检测到致病微生物。我们已经开发了用于CT和模拟尿液中NG检测的样本到答案的分子诊断方法。要测试的假设是，分子诊断芯片可以检测和区分微升规模的人类泌尿生殖系统样本中的CT和NG，其特异性和敏感性优于目前可用的POC检测。我们之前已经在试验台上演示过，芯片的组件可以与模拟样品一起工作。我们的目标是开发一种便携的、廉价的分子诊断方法，可用于POC诊断和鉴别CT和NG感染。我们与约翰·霍普金斯大学性传播疾病护理点技术研究中心的夏洛特·盖多斯博士合作，使用之前收集和存储的已被去除身份和良好特征的泌尿生殖器标本进行了一项中等规模的敏感性和特异性研究。我们将开发一种低成本、集成的样本到结果的分子诊断分析系统来检测衣原体和淋病，该系统使用简单、直观的方法检测核酸扩增产物。为了将这项技术应用于临床和现场，我们提出了以下目标：1.为了使我们目前的测试适应多重CT和NG检测，我们将优化CT、NG的样品制备、扩增和检测，并建立竞争性内部对照(CIC)。2.为了能够在PoC进行测试，我们将优化芯片设计和仪器仪表，以集成样本到答案流程。3.为了在临床上验证新的POC分子诊断方法，我们将使用人类泌尿生殖系统样本，确定该综合检测方法在特异性和敏感性方面是否与金标准实时NAATs相当。该项目的最终成果是一种完全集成的、经过临床验证的STI POC测试分子诊断，该诊断已准备好商业化并扩大到预期的临床试验。

DESCRIPTION (provided by applicant): More than 4 million people in the United States are infected with chlamydia (CT) and gonorrhea (NG). These sexually transmitted infections (STIs) have few early symptoms but cause serious complications that range from sterility and infertility to multidrug resistance and increased HIV infection and transmission. Reducing the spread of these STIs will require early diagnosis, and treatment of infected individuals. Success requires the wide availability of low-cost, robust, and easy to use molecular diagnostics for use at the point-of-care (POC) for the earliest detection of the causative organism. We have developed sample-to-answer molecular diagnostic assays for CT and for NG detection in simulated urine. The hypothesis to be tested is that molecular diagnostic chips can detect and differentiate CT and NG in microliter-scale human urogenital samples with specificity and sensitivity better than presently available POC testing. We have previously demonstrated at the bench that the components of the chip work with simulated samples. Our goal is to develop a portable, inexpensive molecular diagnostic that can be used at the POC to diagnose and differentiate CT and NG infections. We have teamed up with Dr. Charlotte Gaydos at Johns Hopkins University Center for Point-of-Care Technologies Research for Sexually Transmitted Diseases to perform an intermediate sized sensitivity and specificity study using previously collected and stored urogenital specimens that have been de-identified and well characterized. We will develop a low-cost, integrated sample-to-result molecular diagnostic assay system to detect chlamydia and gonorrhea that uses a simple, visual means of detecting nucleic acid amplification products. In order to develop this technology for real-world clinical and field use, the following aims are proposed: 1. To adapt our current test to a multiplexed CT and NG assay, we will optimize sample preparation, amplification, and detection of CT, NG, and a competitive internal control (CIC). 2. To enable testing at the POC, we will optimize the chip design and instrumentation to integrate sample-to-answer processes. 3. To clinically validate the new POC molecular diagnostic, we will determine if the integrated assay is comparable in specificity and sensitivity to gold standard real- time NAATs using human urogenital samples. The ultimate deliverable of the project is a fully-integrated, clinically validated molecular diagnostic for POC testing of STI that is ready for commercialization and scale- up to prospective clinical trials.