An automated, scalable, and rapid AST system for Gonorrhea STIs

淋病是由淋球菌(NG)引起的性传播感染(STI) 感染人类的泌尿生殖器、肛门直肠和咽部。2018年，估计有114万人 美国有病例，全球有8690万病例。令人担忧的是，抗菌素耐药性的发生率不断上升 不仅是由于抗生素的不当使用，还因为NG与基因相互作用并获得了 来自它感染的解剖部位的其他混合感染的物质。如此大范围的抵抗 NG分离株可能蕴藏的机制使开发快速分子 药敏试验(AST)方法。即使在表型AST方法中，到目前为止还没有 被认为对NG执行基于自动肉汤微量稀释的AST是可行的：主要是因为它 在液体培养中是一种挑剔的有机体。因此，用于为NG执行AST的主要方法是 人工琼脂稀释法，这种方法(因为它是如此劳动密集型)目前只在一个非常 公共卫生实验室数量有限。即使是这些实验室，每个月也只检测200-300个样本。 Acenxion生物系统公司提议开发第一个全自动化的NG表型AST平台，该平台既 满足当前疾病预防控制中心的监测指南，并满足未满足的临床需求--AST引导的“定向” 抗生素治疗。我们的仪器将提供最低抑菌浓度(MIC)和敏感/ 中值/阻力(S/I/R)解释在&lt；6小时内完成，样品制备最少(&lt；5分钟/次 样品)，价格合理。该仪器将由可堆叠的盒式读卡器单元组成，最多可有8个 微流体盒容量；软件/PC控制器；一次性消耗品(50孔微流控 预先标有候选抗生素的盒式磁带)。NG采用优化的发酵液和快速搅拌 生长，实现自动微量肉汤稀释。在疾控中心的事先资助下，我们已经展示了 通过建立一个部分自动化的系统来进行肉汤微量检测，对所提议的仪器进行概念验证 在单个16孔盒上进行稀释实验，并用它监测3 NG的生长(或缺失) 菌株(1株ATTC QC株和2株AR分离株)在多种候选抗生素存在的情况下 100%灵敏度和特异度。我们寻求“直接进入第二阶段”的资金来扩展这项早期的工作：建立一个全面的 自动仪器，并与疾病预防控制中心的挑战AR银行小组进行测试，该小组由50种临床分离株组成(跨越 多重抗性机制和产生广泛的表型反应)。我们选择了这套 用微生物代替来自2-3个地点的患者样本，以确保在现实世界中有更好的成功机会。 我们的商业化战略旨在首先为现有市场带来效率(区域实验室参与 在淋球菌分离调查计划中)，然后将用户(客户)基础扩展到医院实验室， 临床实验室网络和临床实验室通过启用AST引导的淋病定向抗生素治疗。

Gonorrhea is a sexually transmitted infection (STI) caused by the bacterium Neisseria gonorrhoeae (NG) that infects the urogenital, anorectal and pharyngeal tracts in humans. In 2018, there were an estimated 1.14M cases in the US, and 86.9M cases worldwide. Of concern is the growing incidence of antimicrobial resistance not only due to inappropriate use of antibiotics, but also because NG interacts with and acquires genetic material from other co-infections in the anatomical sites that it infects. This wide range of resistance mechanisms that NG isolates can potentially harbor makes it very challenging to develop rapid molecular methods of Antibiotic Susceptibility Testing (AST). Even among phenotypic AST methods, it has so far not been considered feasible to perform automated broth microdilution based ASTs on NG: principally because it is a fastidious organism in liquid culture. As a result, the primary method used to perform ASTs for NG is the manual agar dilution method, which (because it is so labor intensive) is currently conducted in only a very limited number of public health labs. Even these labs test only 200-300 samples a month. Acenxion Biosystems proposes to develop the first fully automated NG phenotypic AST platform that both satisfies the current CDC guidance for surveillance and fulfils an unmet clinical need for AST-guided “directed” antibiotic therapy. Our instrument will provide Minimum Inhibitory Concentration (MIC) and Susceptible/ Intermediate /Resistance (S/I/R) interpretation in < 6 hours with minimal sample preparation (<5 min per sample) at a reasonable price. The instrument will consist of stackable, cassette reader units with up to eight microfluidic cassette capacity; software/PC Controller; and single-use consumable (50-well microfluidic cassette pre-spotted with candidate antibiotics). Optimized broth and rapid stirring are employed for NG growth, enabling automated broth microdilution. With prior funding from the CDC, we have demonstrated proof-of-concept for the proposed instrument by building a partly-automated system to perform broth micro- dilution experiments on a single 16-well cassette, and used it monitor the growth (or lack thereof) of 3 NG strains (1 ATTC QC strain, and 2 AR isolate strains) in the presence of multiple candidate antibiotics with 100% sensitivity & specificity. We seek “Direct to Phase II” funds to expand this earlier work: build a fully automated instrument and test it against the CDC’s challenge AR bank panel of 50 clinical isolates (which span multiple resistance mechanisms and yield wide range of phenotypic responses). We chose this set of organisms in lieu of patient samples from 2 - 3 locations to ensure better chances of success in the real world. Our commercialization strategy aims to first bring efficiencies to the existing market (regional labs participating in the Gonococcal Isolate Survey Program) and then to expand the user (customer) base to hospital labs, clinical laboratory networks and clinical labs by enabling AST-guided directed antibiotic therapy for gonorrhea.