SBIR Phase I: Culture-independent Rapid Bacterial Isolation and Identification

这个SBIR一期项目旨在开发和证明一种系统的可行性，该系统可以在1小时内直接从血液中识别出广泛的细菌。现有的诊断系统需要2-6天来识别细菌，这延误了使用适当抗生素的治疗。脓毒症通常是由细菌感染引起的，在美国，高达一半的医院死亡是由延误或治疗不当引起的并发症造成的（这主要是因为缺乏有关引起感染的细菌的信息）。它是美国最贵的住院治疗疾病。相当于203亿美元的医疗费用。该系统旨在大幅减少医生提供所需信息的时间，使他们能够迅速开始使用适当的抗生素进行治疗。预计这将显著降低成本、并发症和死亡率。此外，通过消除培养的需要，可以获得对细菌行为的新见解，有助于对细菌的额外基础知识。这些知识将有助于抗生素发现工作，提高对耐药性机制的理解，通过更快地检测药品和食品的细菌污染以及微生物组的表征，支持新的生物制造工艺。快速细菌鉴定的挑战主要在两个方面？样品制备，消除了培养和多重鉴定的需要，以涵盖能够引起感染的细菌种类的不同范围。提出的工作是基于利用细菌和样品基质中细胞之间的差异反应来选择性地分解样品基质，而不是细菌，这样最大的裂解碎片比完整的细菌小。随后，使用基于尺寸的技术将碎片从完整的细菌中分离出来，从而可以直接从样品中快速分离出广泛的细菌。然后使用新型傅里叶变换红外微光谱系统鉴定临床相关浓度的分离细菌。不使用细菌特异性试剂。建议的第一阶段工作解决了该过程中风险最高的技术障碍。直接从血液中分离和检测低浓度细菌是通过本项目的三个目标来实现的，即相对于传统的基于培养的方法，优化了样品的纯化和细菌回收率、鉴定准确性和集成系统的性能。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。

This SBIR Phase I project aims to develop and demonstrate the feasibility of a system that can identify a broad range of bacteria in 1 hour directly from blood. Existing diagnostic systems require 2-6 days to identify the bacteria, which delays treatment using appropriate antibiotics. Sepsis, which is often caused by a bacterial infection, contributes to up to half of all hospital deaths in the US largely due to complications arising from delays or inappropriate treatment (which is mainly because of lack of information regarding the infection-causing bacteria). It is the most expensive hospital-treated condition in the United States ? representing $20.3 billion in healthcare costs. The proposed system aims to sharply reduce the time to provide information needed by physicians, enabling them to initiate treatment using the appropriate antibiotic rapidly. This is expected to lead to significant reductions in costs, complications, and mortality. Further, by eliminating the need for culturing, new insights into bacterial behavior can be acquired contributing to additional fundamental knowledge of bacteria. This knowledge will aid antibiotic discovery efforts, improve understanding of mechanisms of drug resistance, support new biomanufacturing processes through faster detection of bacterial contamination of pharmaceutical products and foods, and characterization of the microbiome.The challenges of rapid bacterial identification are primarily in two areas ? sample preparation, that eliminates the need for culturing, and multiplex identification, to cover the diverse range of bacterial species capable of causing infection. The proposed work is based on leveraging the differential response between bacteria and the cells in a sample matrix to selectively break down the sample matrix, but not bacteria, such that the largest lysis debris is smaller than intact bacteria. The debris is subsequently separated from intact bacteria using a size-based technique enabling rapid isolation of a broad range of bacteria directly from the sample. A novel Fourier-transform infrared microspectroscopy system is then used to identify the isolated bacteria at clinically relevant concentrations. No bacteria-specific reagents are employed. The proposed Phase I work addresses the highest risk technical hurdles in the process. Isolating and detecting low concentration of bacteria directly from blood is accomplished through the three objectives of this project that optimize the purification and bacterial recovery from the sample, identification accuracy, and performance of the integrated system relative to the traditional culturing-based method.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.