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

小型企业创新研究(SBIR)第一阶段项目的更广泛影响/商业潜力将是开发新的诊断工具，允许在临床环境中进行快速、直接来自样本的血流感染测试。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，同时提供有关抗生素和有效治疗这些感染的剂量的重要表型数据。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，同时还通过减少这些药物的广泛处方来帮助抗击日益增长的多重耐药感染的发生率，这对努力保持现有抗生素的有效性至关重要。拟议的项目将开发一种新的检测方法，用于直接从标本鉴定(ID)、抗菌素敏感性测试(AST)和测定血液中对大肠杆菌的最低抑菌药物浓度(MIC)。现有护理标准的一个重要限制是取得可操作结果所需的时间；目前的方法需要18-72小时。由于所需的时间，医生必须做出经验性的抗菌治疗决定，这往往会导致患者的负面结果。由于细菌用来进化耐药性的不同机制，基因分型方法受到限制，这可能导致假阴性结果。质谱学需要菌落分离，并且只提供ID。然后需要对AST和MIC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，将使用高度特异性的噬菌体混合物，这些噬菌体混合物显示了从组成噬菌体到宿主范围的重叠，快速产生可测量的信号，并且非常适合简单的自动化96孔板免疫分析。该项目的目标是：1)开发一种使用噬菌体特异性抗体的ID和AST/MIC快速免疫分析方法，以及2)证明直接从血样中进行检测的可行性。还将与当前的护理标准诊断策略进行并列比较。预期的结果包括ID、AST和定量MIC在不到5小时内直接在血液中。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.