Metabolic Assessment of Anti-Microbial Susceptibility within One Cell Cycle

项目摘要 越来越多的抗菌素细菌/真菌的出现已经成为一个日益严重的全球威胁 滥用和过度使用抗菌药物。为了对抗感染，减少抗菌素 对于耐药性，在早期检测和表征细菌/真菌对抗生素的敏感性是至关重要的 减少不适当使用抗菌药物和死亡率的感染阶段。要解决这个问题 在紧急的医疗条件下，快速准确地测定沙门氏菌的药物敏感性是至关重要的 细菌/真菌，以便在疾病过程的早期开出最佳治疗药物。传统型 药敏试验方法，如琼脂平板和肉汤稀释法，检测表型 耐药性基于细菌/真菌在抗菌药存在的情况下的生长情况。一位少校 这些方法的局限性在于，它们是以培养为基础的，至少需要16至24小时才能进行。至 针对这一未得到满足的需求，提出了一种微秒级受激拉曼光谱成像平台 能够在复杂环境中以亚微米级的分辨率和早期原位检测单个细菌 测定其对一种抗微生物药物的反应。一个跨学科团队将进行这项拟议的研究。 郑继新博士是相干拉曼散射显微技术的发明者和领先专家。穆罕默德博士 Seleem(co-Pi)是一名DVM科学家，在传染病和微生物学方面拥有广泛的专业知识。瑞安·F·沃克博士 雷里奇(顾问)，印第安纳大学卫生临床病毒学和血清学实验室医学主任， 在传染病的临床诊断方面有丰富的经验。该团队的核心假设是 微秒级相干拉曼光谱成像将使单个微生物细胞的原位分析成为可能 直接从临床样本(全血)中进行浓缩。为了检验这一假设，研究小组将很快证明 微秒尺度新陈代谢受激拉曼成像测定抗菌效果 在单个活细菌中的活性(目标1)，开发微秒尺度的宽带受激拉曼 细菌无标记鉴别和抗微生物检测的光谱显微镜 敏感性(目标2)，并证明真菌的早期检测和快速抗菌药敏感性分析 感染(目标3)。提出的快速AST方法适用于复杂环境中的细菌/真菌和 单细胞水平。因此，长期标本培养和传代培养可获得细菌/真菌分离物。 避免了。与当前的方法相比，这种方法的特点提供了显著的进步 细菌/真菌感染的治疗。

Project Summary The increasing emergence of antimicrobial-resistant bacteria/fungi has become a growing global threat due to the misuse and overuse of antimicrobial drugs. In order to combat infections and reduce anti-microbial resistance, it is essential to detect and characterize bacterial/fungal susceptibility to antimicrobial in the early stages of infections to reduce the inappropriate use of antimicrobial drugs and the death rate. To address this urgent medical condition, it is critical to rapidly and accurately determine the antimicrobial susceptibility of bacteria/fungi so that optimal therapy drugs can be prescribed early in the disease process. Conventional methods for antimicrobial susceptibility testing, such as agar plates and broth dilution assays, detect phenotypic resistance based on bacterial/fungal growth in the presence of antimicrobial drugs being tested. A major limitation of these methods is that they are based on culture and require at least 16 to 24 h to conduct. To address this unmet need, a microsecond-scale stimulated Raman spectroscopic imaging platform is proposed to enable in situ detection of a single bacterium in complex environment at sub-micron resolution and early determination of its response to an antimicrobial drug. An interdisciplinary team will conduct the proposed study. Dr. Ji-Xin Cheng (PI) is an inventor and leading expert in coherent Raman scattering microscopy. Dr. Mohamed Seleem (co-PI) is a DVM-scientist with broad expertise in infectious diseases and microbiology. Dr. Ryan F. Relich (consultant), Medical Director of the Indiana University Health Clinical Virology and Serology Laboratories, has extensive experience in clinical diagnosis of infectious diseases. The team’s central hypothesis that microsecond-scale coherent Raman spectroscopic imaging will enable in situ analysis of single microbial cells enriched directly from a clinical sample (whole blood). To test this hypothesis, the team will demonstrate fast determination of antimicrobial response through microsecond-scale stimulated Raman imaging of metabolic activity in a single living bacterium (aim 1), develop a microsecond-scale broadband stimulated Raman spectroscopic microscope for label-free discrimination of bacteria and determination of anti-microbial susceptibility (aim 2), and demonstrate early detection and fast antimicrobial susceptibility profiling of fungal infections (aim 3). The proposed rapid AST method works for bacteria/fungi in complex environment and at the single cell level. Therefore, long-time specimen culture and subculture to get bacterial/fungal isolate can be avoided. The characteristics of this approach offer a significant advancement over current approaches for treatment of bacterial/fungal infections.