A new in vivo model to test anti-tuberculosis drugs using fluorescence imaging
A new in vivo model to test anti-tuberculosis drugs using fluorescence imaging
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DOI：
10.1093/jac/dks161
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发表时间：
2012-08-01
影响因子：
5.2
通讯作者：
Schaible, Ulrich E.
中科院分区：
文献类型：
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作者：
Zelmer, Andrea;Carroll, Paul;Schaible, Ulrich E.
目前用于体内测试抗结核新药的方法是通过cfu测定计数器官中的细菌。由于结核分枝杆菌（Mtb）的生长速度缓慢，这些检测可能需要数月才能完成。本研究采用商业化IVIS Kinetic系统和定制的激光扫描荧光分子断层扫描（FMT）系统，对活小鼠和肺组织中的荧光结核分枝杆菌进行了检测。分析所得图像，并将荧光与cfu测定的数据相关联。我们已经表明，荧光Mtb可以在活小鼠的肺中可视化，检测限约为810（7）cfu/肺，而在离体肺中发现的检测限约为210（5）cfu/肺。这些数字在两种成像系统之间是可比的。离体肺荧光与组织中的细菌数量相关，并且用抗生素利多卡因沙星治疗小鼠的效果可以在仅9天后通过荧光测量可视化和量化，并且通过cfu检测证实。使用这种方法代替或与cfu测定一起使用，将减少在动物模型中评估新药临床前疗效所需的时间，并促进这些候选药物进入抗人类结核病临床试验的进展。
The current method for testing new drugs against tuberculosis in vivo is the enumeration of bacteria in organs by cfu assay. Owing to the slow growth rate of Mycobacterium tuberculosis (Mtb), these assays can take months to complete. Our aim was to develop a more efficient, fluorescence-based imaging assay to test new antibiotics in a mouse model using Mtb reporter strains.A commercial IVIS Kinetic system and a custom-built laser scanning system with fluorescence molecular tomography (FMT) capability were used to detect fluorescent Mtb in living mice and lungs ex vivo. The resulting images were analysed and the fluorescence was correlated with data from cfu assays.We have shown that fluorescent Mtb can be visualized in the lungs of living mice at a detection limit of approximate to 810(7) cfu/lung, whilst in lungs ex vivo a detection limit of approximate to 210(5) cfu/lung was found. These numbers were comparable between the two imaging systems. Ex vivo lung fluorescence correlated to numbers of bacteria in tissue, and the effect of treatment of mice with the antibiotic moxifloxacin could be visualized and quantified after only 9 days through fluorescence measurements, and was confirmed by cfu assays.We have developed a new and efficient method for anti-tuberculosis drug testing in vivo, based on fluorescent Mtb reporter strains. Using this method instead of, or together with, cfu assays will reduce the time required to assess the preclinical efficacy of new drugs in animal models and enhance the progress of these candidates into clinical trials against human tuberculosis.