Rapid and Cost-Effective Diagnostic System for Sexually Transmitted Infections

描述（由申请人提供）：根据CDC，衣原体和淋病分别由沙眼衣原体（CT）和淋病奈瑟菌（NG）引起，是美国和世界其他地区报告的第一和第二常见性传播感染。这些性传播感染往往诊断不足，导致治疗延迟，持续传播和公共医疗费用增加（每年约147亿美元）。由于性传播感染的流行和长期健康后果，需要快速、敏感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，因为他们很少返回第二次就诊。护理点（POC）测试是应对性传播感染流行病的一项重要战略，由于许多受性传播感染的人（年轻人和/或穷人）通常无法获得传统医疗保健，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POC测试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CT和NG等性病病原体。该系统将结合联合收割机Lynntech的新型核酸扩增和侧流检测设备，可用于许多非传统的医疗保健环境。在第一阶段，将开发高性能、低成本的热梯度循环仪（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反应（PCR），该架构从单一热源产生稳定的热梯度。它解决了许多热循环仪常见的问题，如用户界面，样品加载，可靠性和再现性。TGC将与用于检测PCR扩增子的侧向流条带组件偶联，并将该组件并入标准0.2 mL PCR管中，该PCR管在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，并且它允许非技术用户在没有样品转移或特殊仪器的情况下轻松读取和解释结果。将证明整个试验可在45 min内完成，包括样品制备。当与适当的样品制备模块（也由Lynntech开发）结合时，该系统可以作为快速诊断STI的POC设备。拟议的工作将与来自各个领域的专家合作进行，包括夏洛特盖多斯（STI专家，约翰霍普金斯大学）、刘国栋（纳米材料和侧向流分析，北达科他州州立大学）、维克托乌加兹（流体/热表征，得克萨斯A&M大学）和唐毅伟（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，因为他们在侧流检测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如何在POC或非传统医疗环境中实施复杂的分子诊断检测。 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），导致严重的长期健康问题，如败血症，狭窄疾病，盆腔炎（PID），异位妊娠和不孕症。该项目将推进POC分子诊断设备的开发，用于在非传统医疗环境中快速和廉价地筛查性传播感染。Lynntech将建立一个非常低的成本（75美元），自动热梯度循环仪（TGC），以执行多达16个聚合酶链反应，在不到25分钟。此外，Lynntech将结合联合收割机这种TGC技术与横向流动测试技术的PCR扩增子检测。这种低成本（<2美元）、5分钟侧向流动的一次性装置专门设计用于进行“闭管”分子检测。如果与样本制备方法适当结合，这两项创新技术将产生45分钟的分子检测，以识别性传播感染病原体，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.