A smartphone-based point-of-care test for detection of Neisseria gonorrhoeae

摘要 该项目的最终目标是CLIP-G，这是一项FDA批准和CE认证的基于智能手机的测试（≤ 30分钟） 用于女性和男性的淋病奈瑟菌（NG）检测，用于非处方药（OTC）和点 护理（POC）设置。NG导致第二大常见的细菌性性传播感染（STI）， 美国每年新增55万例。未经治疗的人有很高的风险，严重的后遗症，包括骨盆 炎症性疾病、异位妊娠和不孕症。大多数NG感染可以用抗生素治愈，但 事实上，它们在大多数感染者中不引起症状或非特异性症状， 测试方法目前推荐的核酸扩增测试（NAAT）是昂贵的，并且 缓慢的周转时间;这限制了它们扩展测试覆盖范围的适用性。快速POC测试可以抑制 通过在一个诊所中启用"检测和治疗"方法并立即通知合作伙伴来传播疾病 访问也可用于POC设置的OTC测试将扩展测试到不需要的风险个体 由于性传播感染的耻辱感，目前正在寻求检测。CLIP-G使用"纳米荧光粉"-持久发光 无机纳米颗粒-作为侧流测定（LFA）报告物用于NG特异性抗原的免疫测定， 结合智能手机的闪光灯和摄像头以及一个便宜的读出适配器。CLIP平台 与传统的LFA相比，能够实现数量级更好的检测限。当前基于抗原的测定 对于NG，它们是高度特异性的，但由于它们的高检测限（LOD）而具有差的临床灵敏度。中央 该项目的假设是，与现有NG相比，CLIP-G免疫测定的LOD大大改善 与NAAT相比，免疫测定将导致> 90%的临床灵敏度，而不损害高灵敏度。 特异性> 98%。在这个快速通道项目的第一阶段，我们将开发"v1 CLIP-G"台式原型， 敏感性和特异性NG检测，并使用传代培养的临床NG评价初步分析包容性 我们还将在一项小N回顾性临床试验中评估v1 CLIP-G原型的临床可行性。 study.假设我们可以在第一阶段获得该项目中心假设的初步验证，我们将采取行动， 进入第二阶段，在第二阶段中，我们将首先迭代地设计和开发辅助硬件和软件组件 完全集成的CLIP-G试剂盒。由此开发的CLIP-G套件将 在可用性研究、综合分析性能研究和大型（n> 1，000）临床研究中进行了评估 使用前瞻性收集的样本进行研究。Luminostics已经组建了一个由生物工程师，化学家， 软件工程师，微生物学家，临床医生和制造专家，以最大限度地提高成功的几率。的 该快速通道项目的预期结果是显著降低针对FDA 510（k）的最终试验的风险 CLIP-G的OTC许可和纳米磷-LFA作为平台技术的进一步验证， 在各种医疗条件下的潜在应用，可以使用更敏感的POC测试。

ABSTRACT The ultimate goal of this project is CLIP-G, an FDA-cleared and CE-marked smartphone-based test (≤30 min) for Neisseria gonorrhoeae (NG) detection in women and men for use in over-the-counter (OTC) and point-of- care (POC) settings. NG causes the 2nd-most common bacterial sexually transmitted infection (STI) with over 550,000 new cases in the US annually. Untreated individuals are at high risk of serious sequelae including pelvic inflammatory disease, ectopic pregnancy, and infertility. Most NG infections are curable with antibiotics, but the fact that they cause no symptoms or non-specific symptoms in a majority of those infected necessitates accurate testing methods. Currently recommended Nucleic Acid Amplification Tests (NAATs) are expensive and have slow turnaround times; this limits their applicability for expanding testing coverage. Rapid POC tests can curb disease transmission by enabling a "test and treat" approach and immediate partner notification in a single clinic visit. An OTC test that can also be used in POC settings will expand testing to the at-risk individuals who do not currently seek testing due to the stigma around STIs. CLIP-G uses "nanophosphors" – persistent luminescent inorganic nanoparticles – as lateral flow assay (LFA) reporters in an immunoassay for NG-specific antigens in combination with a smartphone’s flash and camera and an inexpensive adapter for readout. The CLIP platform enables orders-of-magnitude better detection limits compared to traditional LFAs. Current antigen-based assays for NG are highly specific but have poor clinical sensitivity due to their high limit of detection (LOD). The central hypothesis of this project is that the vastly improved LOD of the CLIP-G immunoassay compared to existing NG immunoassays will result in clinical sensitivities >90% compared to NAATs without compromising on high specificity (>98%). In Phase I of this Fast-Track project, we will develop “v1 CLIP-G” benchtop prototypes for sensitive and specific NG detection and evaluate preliminary analytical inclusivity using sub-cultured clinical NG isolates; we will also evaluate the clinical feasibility of v1 CLIP-G prototypes in a small-N retrospective clinical study. Assuming we can obtain preliminary validation of the project’s central hypothesis in Phase I, we will move on to Phase II, in which we’ll first iteratively design and develop the ancillary hardware and software components of the fully integrated CLIP-G kit around the Phase I-developed assay. Thus-developed CLIP-G kits will be evaluated in usability studies, comprehensive analytical performance studies, and a large (n >1,000) clinical study using prospectively collected specimens. Luminostics has assembled a team of bioengineers, chemists, software engineers, microbiologists, clinicians, and manufacturing experts to maximize the odds of success. The expected outcome of this Fast-Track project is the significant de-risking of the final trial targeting FDA 510(k) OTC clearance for CLIP-G and the further validation of nanophosphor-LFAs as a platform technology with potential applications in a variety of medical conditions that could use more sensitive POC tests.