Rapid and Cost-Effective Diagnostic System for Sexually Transmitted Infections

描述（由申请人提供）：根据CDC，分别由沙眼衣原体（CT）和淋病奈瑟菌（NG）引起的衣原体和淋病是美国和世界其他地区报告的第一和第二大最常见的性传播感染。这些性传播感染往往未得到充分诊断，导致治疗延误、持续传播和更高的公共医疗费用（每年约147亿美元）。由于性传播感染的普遍性和对健康的长期影响，需要有快速、灵敏的方法来检测性传播感染，以便患者立即得到结果和治疗。重要的是，患者在第一次就诊时获得检测结果和初步咨询或治疗，因为他们很少再次就诊。即时检测是应对性传播感染流行的一项重要战略，由于许多受性传播感染影响的人（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、社区公共卫生保健诊所、监狱、拘留中心）进行即时检测，可以大大减少性传播感染。该提案的目的是开发一种快速、易于使用、廉价的诊断系统，用于识别诸如CT和NG等STI病原体。该系统将结合Lynntech的新型核酸扩增和侧流检测设备，可用于许多非传统医疗保健环境。在第一阶段，将开发一种高性能、低成本的热梯度循环器（TGC）。它将在简化的结构中对CT/NG目标进行快速聚合酶链反应（PCR），从单一热源产生稳定的热梯度。它解决了许多热循环器常见的问题，例如用户界面，样品加载，可靠性和可重复性。TGC将与用于检测PCR扩增子的横向流动条带组件耦合，该组件被纳入标准的0.2 ml PCR管中，在PCR期间用于TGC。这种优雅的集成允许在封闭系统中进行扩增和检测，消除PCR后污染，并允许非技术用户轻松读取和解释结果，无需样品转移或特殊仪器。将证明整个分析可以在不到45分钟内完成，包括样品制备。当与合适的样品制备模块（也由Lynntech开发）结合使用时，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各个领域的专家合作进行，包括Charlotte Gaydos （STI专家，约翰霍普金斯大学），刘国栋（纳米材料和横向流动试验，北达科他州立大学），Victor Ugaz（流体/热表征，德克萨斯A&M大学）和Tang i- wei（临床样品制备，范德比尔特大学）。为了协助技术商业化，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具有专业知识。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。

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.