Direct continuous method for monitoring biofilm infection in a mouse model
Direct continuous method for monitoring biofilm infection in a mouse model
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DOI：
10.1128/iai.71.2.882-890.2003
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发表时间：
2003-02-01
影响因子：
3.1
通讯作者：
Contag, PR
中科院分区：
文献类型：
--
作者：
Kadurugamuwa, JL;Sin, L;Contag, PR
我们开发了一种快速、连续的方法，通过对聚四氟乙烯导管上定植的生物发光细菌进行无创成像，在体外和小鼠感染模型中实时监测生物膜。两种重要的生物膜形成细菌病原体，金黄色葡萄球菌和铜绿假单胞菌，通过插入完整的 lux 操纵子而产生生物发光。这些细菌为体外研究和体内模型的开发产生了显着的生物发光信号，从而可以有效地实时评估生物膜的生理状态。体外活菌计数和光输出是平行且高度相关的（金黄色葡萄球菌 r = 0.98；铜绿假单胞菌 r = 0.99），并且可以维持 10 天或更长时间，前提是每 12 It 补充一次生长培养基。在小鼠模型中，皮下植入导管（预定植或植入后感染）具有良好的耐受性。金黄色葡萄球菌和铜绿假单胞菌的感染剂量为 10 (3) 至 10 (5) CFU/导管，导致导管周围出现可重复的局部感染，这种感染一直持续到第 20 天实验结束。从受感染动物的导管中回收细菌表明，生物发光信号与 CFU 相对应，并且即使在体内许多天后，lux 构建体也高度稳定。由于可以直接在支持基质上非破坏性、非侵入性地检测活细胞的代谢活性，因此该方法对于慢性生物膜感染的研究和体内药效研究特别有吸引力。
We have developed a rapid, continuous method for real-time monitoring of biofilms, both in vitro and in a mouse infection model, through noninvasive imaging of bioluminescent bacteria colonized on Teflon catheters. Two important biofilm-forming bacterial pathogens, Staphylococcus aureus and Pseudomonas aeruginosa, were made bioluminescent by insertion of a complete lux operon. These bacteria produced significant bioluminescent signals for both in vitro studies and the development of an in vivo model, allowing effective real-time assessment of the physiological state of the biofilms. In vitro viable counts and light output were parallel and highly correlated (S. aureus r = 0.98; P. aeruginosa r = 0.99) and could be maintained for 10 days or longer, provided that growth medium was replenished every 12 It. In the murine model, subcutaneous implantation of the catheters (precolonized or postimplant infected) was well tolerated. An infecting dose of 10 (3) to 10 (5) CFU/catheter for S. aureus and P. aeruginosa resulted in a reproducible, localized infection surrounding the catheter that persisted until the termination of the experiment on day 20. Recovery of the bacteria from the catheters of infected animals showed that the bioluminescent signal corresponded to the CFU and that the lux constructs were highly stable even after many days in vivo. Since the metabolic activity of viable cells could be detected directly on the support matrix, nondestructively, and noninvasively, this method is especially appealing for the study of chronic biofilm infections and drug efficacy studies in vivo.