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

淋病是一种性传播感染（STI），由细菌淋病奈瑟菌（NG）引起， 感染人类的泌尿生殖道、肛门直肠道和咽道。2018年，估计有114万人 美国有8690万例，全球有8690万例。令人担忧的是， 不仅由于抗生素的不适当使用，而且由于NG与遗传因子相互作用并获得遗传因子， 来自其感染的解剖部位中的其他合并感染的材料。这种大范围的抵抗 NG分离株可能潜在的机制使得开发快速分子生物学技术非常具有挑战性。 抗生素敏感性试验（AST）。即使在表型AST方法中，迄今为止也没有 认为对NG进行基于自动肉汤微量稀释的AST是可行的：主要是因为 在液体培养中是一种要求苛刻的微生物。因此，用于执行NG AST的主要方法是 手动琼脂稀释法，其中（因为它是如此劳动密集型）目前仅在一个非常 公共卫生实验室数量有限。即使是这些实验室，每个月也只能检测200-300个样本。 Acenxion Biosystems提议开发第一个全自动NG表型AST平台， 满足了当前CDC的监测指南，并满足了AST指导的“定向”临床需求 抗生素治疗我们的仪器将提供最低抑菌浓度（MIC）和敏感/ 中间/电阻（S/I/R）解释在< 6小时内进行，样品制备最少（每个 样品）以合理的价格。该仪器将由可堆叠的试剂卡读取器单元组成，最多可容纳8个 微流体盒容量;软件/PC控制器;和一次性消耗品（50孔微流体 用候选抗生素预点样的盒）。NG采用优化的发酵液和快速搅拌 生长，实现自动肉汤微量稀释。在疾病预防控制中心的资助下，我们已经证明了 通过建立一个部分自动化的系统来执行肉汤微量测定， 在单个16孔盒上进行稀释实验，并使用它监测3 NG的生长（或缺乏生长 菌株（1株ATTC QC菌株和2株AR分离株）在存在多种候选抗生素的情况下， 100%灵敏度和特异性。我们寻求“直接进入第二阶段”的资金，以扩大这一早期工作：建立一个全面的 自动化仪器，并将其与CDC的50种临床分离株的挑战AR库面板进行测试（该面板跨越 多种抗性机制和产生广泛的表型反应）。我们选择了这套 微生物代替来自2 - 3个地点的患者样本，以确保在真实的世界中更好的成功机会。 我们的商业化战略旨在首先提高现有市场的效率（区域实验室参与 在淋球菌分离株调查计划中），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，使AST指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.