SBIR Phase I: Phage-amplified Identification and Antimicrobial Susceptibility Test

这个小企业创新研究（SBIR）第一阶段项目的更广泛的影响/商业潜力将是开发新的诊断工具，以便在临床环境中快速、直接从样本进行血液感染检测。这里提出的新技术创新将更快地识别导致绝大多数血液感染的细菌，同时提供重要的抗生素表型数据和治疗这些感染的有效剂量。这将通过更快的诊断和更精确的抗生素选择来改善患者的预后，同时还有助于通过减少这些药物的广泛处方来对抗日益增长的耐多药感染发生率，这对保持现有抗生素的有效性至关重要。该项目将开发一种新的检测方法，用于血液中大肠杆菌的直接标本鉴定（ID）、抗菌药物敏感性试验（AST）和最低抑制药物浓度测定（MIC）。现有护理标准的一个重大限制是取得可采取行动的结果需要时间；目前的方法需要18-72小时。由于需要时间，医生必须做出经验性的抗菌治疗决定，这往往导致患者的负面结果。基因型方法是有限的，因为细菌利用不同的机制来进化耐药性，这可能导致假阴性结果。质谱法需要菌落分离，只能提供ID。AST和MIC需要进行其他耗时的测试。需要快速的表型方法。为了解决这些问题，噬菌体混合物将被用于高度特异性的噬菌体混合物，这些噬菌体具有与组成噬菌体重叠的宿主范围，可以快速产生可测量的信号，并且非常适合简单的自动化96孔板免疫测定。项目目标是：1)开发一种使用噬菌体特异性抗体的快速ID和AST/MIC免疫测定方法；2)证明直接从血液样本中检测的可行性。还将与目前的标准诊断策略进行并排比较。预期结果包括5小时内直接在血液中的ID、AST和定量MIC。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project will be the development of new diagnostic tools that allow rapid, direct-from-specimen bloodstream infection testing in clinical settings. The new technological innovations proposed here will more quickly identify bacteria responsible for the vast majority of blood infections while providing vital phenotypic data on antibiotics and dosages effective for treating these infections. This will improve patient outcomes through faster diagnosis and more precise antibiotic selection while also helping to combat the growing incidence of multidrug-resistant infections by reducing broad prescription of these drugs, which is critical to efforts to preserve the usefulness of existing antibiotics. The proposed project will develop a new tests for direct-from-specimen identification (ID), antimicrobial susceptibility testing (AST) and minimum inhibitory drug concentration (MIC) determination against Escherichia coli in blood. A significant limitation with existing standards-of-care is time required for actionable results; current approaches require 18-72 hrs. Due to the time required, physicians must make empiric antimicrobial therapy decisions that often lead to negative patient outcomes. Genotypic approaches are limited because of the varied mechanisms utilized by bacteria to evolve resistance, which can lead to false-negative results. Mass spectrometry requires colony isolation and provides ID only. Other time-consuming tests are then required for AST and MIC. Rapid phenotypic approaches are needed. To solve these problems, bacteriophage mixtures will be used that are highly specific and have demonstrated overlapping host ranges from component phages, produce a measurable signal quickly, and are well-suited to a simple automatable 96-well plate immunoassay. Project objectives of are to: 1) develop a rapid ID and AST/MIC immunoassay using phage-specific antibodies, and 2) prove feasibility of direct-from-specimen testing in blood samples. Side-by-side comparison to the current standard-of-care diagnostic strategies will also be performed. Anticipated results include ID, AST, and quantitative MIC in less than 5 hours directly in blood.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.