Rapid and Cost-Effective Diagnostic System for Sexually Transmitted Infections

描述(申请人提供)：根据疾控中心，沙眼衣原体(CT)和淋病(NG)分别是美国和世界其他地区最常见的性传播感染报告的第一和第二位。这些性传播感染往往被低估，导致治疗延迟、持续传播和更高的公共医疗费用(每年约147亿美元)。由于性传播感染的流行和长期的健康后果，需要快速、灵敏的检测性传播感染的方法，以便患者立即获得结果和治疗。重要的是，患者在第一次就诊期间收到检测结果和初步咨询或治疗，因为他们很少再回来第二次就诊。医疗点(POC)检测是应对STI流行的重要战略，由于许多受STI影响的人(年轻人和/或穷人)获得传统医疗保健的机会往往有限，因此可以在非传统环境(大学诊所、社区公共卫生诊所、监狱、拘留中心)进行的POC检测可以极大地减少STI。这项提议的目的是开发一种快速、易用、廉价的诊断系统，用于识别CT和NG等STI病原体。该系统将结合Lynntech的新型核酸放大和横向流动检测设备，可用于许多非传统医疗环境。在第一阶段，将开发一种高性能、低成本的热梯度循环器(TGC)。它将在一个简化的架构中对CT/NG靶标进行快速聚合酶链式反应(PCR)，从单一热源创建稳定的温度梯度。它解决了许多热循环仪的常见问题，如用户接口、样品加载、可靠性和重复性。TGC将与用于检测PCR扩增的横向流动条组件耦合，并将该组件并入标准的0.2-mL聚合酶链式反应试管中，该试管在聚合酶链式反应过程中用于组氨酸聚合酶链式反应。这种优雅的集成使扩增和检测可以在封闭的系统中进行，消除了聚合酶链式反应后的污染，并允许非技术用户轻松阅读和解释结果，而无需转移样本或使用特殊仪器。这将证明，整个分析可以在不到45分钟内完成，包括样品准备。当与适当的样品制备模块(也由Lynntech开发)相结合时，该系统可以作为快速诊断性传播感染的POC设备。拟议的工作将与来自不同领域的专家合作开展，包括约翰·霍普金斯大学的夏洛特·盖多斯(STI专家)、刘国栋(北达科他州立大学的纳米材料和侧向流动分析)、维克多·乌加兹(得克萨斯农工大学的流体/热表征)和唐一伟(范德比尔特大学的临床样品制备)。为了帮助技术商业化，诊断咨询网络公司将提供咨询，因为他们在横向流动分析设计和制造方面拥有专业知识。我们技术的成功开发和商业化将影响复杂的分子诊断分析在POC或非传统医疗保健环境中的实施方式。 公共卫生相关性：衣原体和淋病是美国最常见的性传播感染(STI)，会导致严重的长期健康问题，如败血症、狭窄疾病、盆腔炎(PID)、宫外孕和不孕。该项目将推动POC分子诊断设备的开发，用于在非传统医疗环境中快速且廉价地筛查性传播感染。Lynntech将制造一种成本非常低(75美元)的自动温度梯度循环仪(TGC)，在不到25分钟的时间内进行多达16次聚合酶链式反应。此外，Lynntech将把这项TGC技术与横向流动测试技术相结合，用于PCR扩增片段的检测。这种低成本(2美元)、5分钟侧向流动的一次性设备是专门为执行“闭管”分子检测而设计的。如果与样品制备方法适当结合，这两项创新技术将产生45分钟的分子测试，以识别STI病原体，以便患者在离开医疗机构之前立即接受治疗。

DESCRIPTION (provided by applicant): According to the CDC, chlamydia and gonorrhea, caused by Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG), respectively, are the first and second most frequently reported STIs in the US and likely the rest of the world. These STIs are often under-diagnosed, leading to delayed treatment, continued spread and higher public healthcare costs (~$14.7 billion annually). As a result of their prevalence and long term health consequences, there is a need for rapid, sensitive methods of detecting STIs in order for patients to get results and treatment immediately. It is important that patients receive test results and initial consultation or treatment during the first patient visit, as they rarely return for a second visit. Point-of-care (POC) tests are an important strategy to address the STI epidemic, and because many affected by STIs (young and/or poor) often have limited access to traditional healthcare, POC tests that can be performed in non-traditional settings (university clinics, community public health care clinics, jails, detention centers) could greatly reduce STIs. The aim of this proposal is to develop a rapid, easy-to-use, inexpensive diagnostic system for identifying STI pathogens such as CT and NG. The system will combine Lynntech's novel nucleic acid amplification and lateral flow detection devices and can be used in many non-traditional healthcare settings. In Phase I, a high performance, low-cost thermal gradient cycler (TGC) will be developed. It will perform rapid polymerase chain reaction (PCR) for CT/NG targets in a simplified architecture that creates a stable thermal gradient from a single heat source. It addresses problems common to many thermal cyclers, such as user interfacing, sample loading, reliability and reproducibility. The TGC will be coupled with a lateral flow strip component for detecting PCR amplicons, and this component is incorporated into a standard 0.2-mL PCR tube, which is used in the TGC during PCR. This elegant integration allows the amplification and detection to be performed in a closed system, eliminating post- PCR contamination, and it permits a non-technical user to easily read and interpret results without sample transfer or special instrumentation. It will be demonstrated that the entire assay can be completed in less than 45 min, including sample preparation. When combined with an appropriate sample preparation module (also being developed by Lynntech), the system could serve as a POC device for rapid diagnosis of STIs. The proposed work will be carried out in collaboration with experts from various fields, including Charlotte Gaydos (STI expert, Johns Hopkins University), GuoDong Liu (nanomaterials and lateral flow assay, North Dakota State University), Victor Ugaz (fluidic/thermal characterization, Texas A&M University), and Yi-Wei Tang (clinical sample preparation, Vanderbilt University). To assist with technology commercialization, Diagnostic Consulting Network, Inc will provide consultation, as they have expertise in lateral flow assay design and manufacturing. The successful development and commercialization of our technology will impact how sophisticated molecular diagnostic assays can be implemented in POC or non-traditional healthcare settings. PUBLIC HEALTH RELEVANCE: Chlamydia and gonorrhea are the most common sexually transmitted infections (STIs) in the US, causing serious long term health problems such as sepsis, stricture disease, pelvic inflammatory disease (PID), ectopic pregnancy and infertility. This project will advance the development of a POC molecular diagnostic device for the rapid and inexpensive screening of STIs in non-traditional healthcare settings. Lynntech will build a very low cost ($75 parts), automated thermal gradient cycler (TGC) to perform up to 16 polymerase chain reactions in less than 25 min. Additionally, Lynntech will combine this TGC technology with a lateral flow test technology for PCR amplicon detection. This low cost (<$2), 5-min lateral flow disposable device is specifically designed to perform "closed-tube" molecular detection. If properly coupled with a sample preparation method, these two innovative technologies will yield a 45-min molecular test to identify STI pathogens in order for patients to receive immediate treatment before they leave a healthcare facility.