Rapid in vivo assessment of drug efficacy against Mycobacterium tuberculosis using an improved firefly luciferase.
Rapid in vivo assessment of drug efficacy against Mycobacterium tuberculosis using an improved firefly luciferase.
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DOI：
10.1093/jac/dkt155
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发表时间：
2013-09
期刊：
影响因子：
--
通讯作者：
Robertson BD
中科院分区：
文献类型：
--
作者：
Andreu N;Zelmer A;Sampson SL;Ikeh M;Bancroft GJ;Schaible UE;Wiles S;Robertson BD
体内实验是昂贵且耗时的，并且呈现出抗结核药物开发的主要瓶颈。常规方法依赖于细菌菌落的计数，结核分枝杆菌在琼脂平板上生长可能需要长达4周的时间。由表达荧光素酶的重组细菌产生的光可以用作细菌负荷的标记，并且通过使用适当的成像设备可以在活动物中容易地非侵入性地跟踪疾病进展。本工作的目的是建立一种基于生物发光的结核病小鼠模型，以评估抗生素对结核分枝杆菌的疗效。体内结核病我们用了M。携带萤火虫荧光素酶基因（FFlucRT）的红移衍生物的结核菌株感染小鼠，并在一线抗结核药物异烟肼治疗前后通过生物发光成像监测活体动物的疾病进展。分析所得图像，并将生物发光与细菌计数相关联。使用生物发光成像，我们在小鼠肺中分别检测到1.7× 103和7.5 ×104个报告细菌。 在这两种情况下，发现生物发光和细菌负荷之间有很好的相关性。此外，在给予异烟肼治疗的活小鼠中观察到发光显著减少。我们已经证明，一个改进的生物发光菌株的M。在急性进行性感染期间，可以通过活小鼠的非侵入性成像来观察结核病，并且该技术可以用于快速观察和量化抗生素治疗的效果。我们相信，在这里提出的模型将是一个很大的好处，在早期的药物发现作为一个简单，快速的方法来确定活性化合物在体内。
In vivo experimentation is costly and time-consuming, and presents a major bottleneck in anti-tuberculosis drug development. Conventional methods rely on the enumeration of bacterial colonies, and it can take up to 4 weeks for Mycobacterium tuberculosis to grow on agar plates. Light produced by recombinant bacteria expressing luciferase enzymes can be used as a marker of bacterial load, and disease progression can be easily followed non-invasively in live animals by using the appropriate imaging equipment. The objective of this work was to develop a bioluminescence-based mouse model of tuberculosis to assess antibiotic efficacy against M. tuberculosis in vivo. We used an M. tuberculosis strain carrying a red-shifted derivative of the firefly luciferase gene (FFlucRT) to infect mice, and monitored disease progression in living animals by bioluminescence imaging before and after treatment with the frontline anti-tuberculosis drug isoniazid. The resulting images were analysed and the bioluminescence was correlated with bacterial counts. Using bioluminescence imaging we detected as few as 1.7 × 103 and 7.5 × 104 reporter bacteria ex vivo and in vivo, respectively, in the lungs of mice. A good correlation was found between bioluminescence and bacterial load in both cases. Furthermore, a marked reduction in luminescence was observed in living mice given isoniazid treatment. We have shown that an improved bioluminescent strain of M. tuberculosis can be visualized by non-invasive imaging in live mice during an acute, progressive infection and that this technique can be used to rapidly visualize and quantify the effect of antibiotic treatment. We believe that the model presented here will be of great benefit in early drug discovery as an easy and rapid way to identify active compounds in vivo.
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影响因子：
11.3
作者：
Andreu N;Zelmer A;Wiles S
通讯作者：
Wiles S
影响因子：
3.7
作者：
Zhang T;Fang Y;Wang X;Deng X;Zhang X;Hu S;Yu J
通讯作者：
Yu J
影响因子：
4.9
作者：
Rullas, Joaquin;Ignacio Garcia, Juan;Angulo-Barturen, Inigo
通讯作者：
Angulo-Barturen, Inigo
DOI：
10.1016/0092-1157(87)90012-6
发表时间：
1987-01-01
期刊：
JOURNAL OF BIOLOGICAL STANDARDIZATION
影响因子：
--
作者：
JANASZEK, W;ALEKSANDROWICZ, J;SITKIEWICZ, D
通讯作者：
SITKIEWICZ, D
DOI：
10.1073/pnas.1000643107
发表时间：
2010-07-06
影响因子：
11.1
作者：
Kong, Ying;Yao, Hequan;Cirillo, Jeffrey D.
通讯作者：
Cirillo, Jeffrey D.