Validation of a noninvasive, real-time imaging technology using bioluminescent Escherichia coli in the neutropenic mouse thigh model of infection
Validation of a noninvasive, real-time imaging technology using bioluminescent Escherichia coli in the neutropenic mouse thigh model of infection
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DOI：
10.1128/aac.45.1.129-137.2001
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发表时间：
2001-01-01
影响因子：
4.9
通讯作者：
Parr, TR
中科院分区：
文献类型：
--
作者：
Rocchetta, HL;Boylan, CJ;Parr, TR
验证了一种非侵入性、实时检测技术在定性和定量抗菌治疗中的应用。利用多拷贝质粒将发光光habdusluminesens的lax基因簇导入大肠杆菌临床分离株EC14。采用中性粒细胞减少小鼠大腿感染模型，研究该生物发光报告菌在体外和体内的抗菌作用。利用强化电荷耦合器件（ICCD）相机系统监测和测量体外和体内的生物发光，并将这些结果与使用常规平板计数方法测定的活细胞结果进行比较。统计分析表明，在存在或不存在抗菌药物（头孢他啶、四环素或环丙沙星）的情况下，体外和体内生物发光水平与活细胞计数之间存在很强的相关性。使用任何一种方法都可以可靠地进行体内抗菌药物的评估，因为每种方法都是治疗成功的良好指标。剂量依赖性反应也可以通过使用生物发光或活细胞计数作为标记，在中性粒细胞减少小鼠大腿模型中检测到。此外，还研究了ICCD技术在治疗研究期间反复监测同一动物的益处。重复测量同一动物的能力减少了治疗实验中的可变性，并允许在确定治疗效果方面具有相同或更大的信心。这项技术可以减少在此类研究中使用的动物数量，并在药物发现过程中对测试化合物进行评估。
A noninvasive, real-time detection technology was validated for qualitative and quantitative antimicrobial treatment applications. The lax gene cluster of Photorhabdus luminescens was introduced into an Escherichia coli clinical isolate, EC14, on a multicopy plasmid. This bioluminescent reporter bacterium was used to study antimicrobial effects in vitro and in vivo, using the neutropenic-mouse thigh model of infection. Bioluminescence was monitored and measured in vitro and in vivo with an intensified charge-coupled device (ICCD) camera system, and these results were compared to viable-cell determinations made using conventional plate counting methods. Statistical analysis demonstrated that in the presence or absence of antimicrobial agents (ceftazidime, tetracycline, or ciprofloxacin), a strong correlation existed between bioluminescence levels and viable cell counts in vitro and in vivo. Evaluation of antimicrobial agents in vivo could be reliably performed with either method, as each was a sound indicator of therapeutic success. Dose-dependent responses could also be, detected in the neutropenic-mouse thigh model by using either bioluminescence or viable-cell counts as a marker. In addition, the ICCD technology was examined for the benefits of repeatedly monitoring the same animal during treatment studies. The ability to repeatedly measure the same animals reduced variability within the treatment experiments and allowed equal or greater confidence in determining treatment efficacy. This technology could reduce the number of animals used during such studies and has applications for the evaluation of test compounds during drug discovery.