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目的建立产气荚膜梭菌感染小鼠的后肢的气性坏疽动物模型,为早期诊断、了解气性坏疽的致病机理及治疗方法提供依据。方法小鼠随机分成4组,每组10只。三个实验组分别肌肉注射3.5×109、3.5×108和3.5×107cfu/ml浓度的产气荚膜梭菌(ATCC13124)菌液0.1ml,空白对照组肌肉注射生理盐水0.1ml,72h后观察小鼠感染情况,取伤口分泌物作革兰氏染色涂片,细菌血平板厌氧培养和荧光定量PCR方法定量检测。结果 3.5×109、3.5×108、3.5×107cfu/ml实验组和对照组在肌肉注射72h内的死亡率为90%、70%、10%和0%。各组平均存活时间依次为(20:43±11:12)h、(37:24±25:39)h、(68:36±10:45)h和(72:00±0:00)h,死亡小鼠出现了气性坏疽的症状,分泌物培养和镜检出产气荚膜梭菌;未死亡小鼠康复;空白对照组无任何症状。各组Ct值均值为21.21±2.69、28.45±2.74、32.49±2.87和0.00±0.00,组间P值均<0.05。结论成功建立了不同浓度的产气荚膜梭菌在不同时间段内感染小鼠的后肢气性坏疽的动物模型,为了解气性坏疽的致病机理及治疗方法奠定基础。
Objective To establish an animal model of gas gangrene in the hindlimb of Clostridium perfringens infected mice and provide evidence for early diagnosis and understanding of the pathogenesis and treatment of gas gangrene. Methods Mice were randomly divided into 4 groups with 10 mice in each group. 0.1 ml of C. perfringens (ATCC13124) was intramuscularly injected into the three experimental groups at a dose of 3.5 × 109, 3.5 × 108 and 3.5 × 107 cfu / ml, respectively. The control group was intramuscularly injected with 0.1 ml of physiological saline and observed after 72 hours Rat infection, take wounds secretions Gram stain smear, bacterial blood plate anaerobic culture and quantitative PCR quantitative detection. Results The mortality rates of experimental group and control group were 90%, 70%, 10% and 0% within 72 hours after intramuscular injection respectively at 3.5 × 109, 3.5 × 108 and 3.5 × 107 cfu / ml. The mean survival time in each group was (20:43 ± 11: 12) h, (37:24 ± 25:39) h, (68:36 ± 10:45) h and (72:00 ± 0:00) h , The symptoms of gas gangrene appeared in the dead mice, Clostridium perfringens cultured in secretions and microscopically detected, the mice survived without any recovery, and the control group without any symptoms. The average value of Ct in each group was 21.21 ± 2.69,28.45 ± 2.74,32.49 ± 2.87 and 0.00 ± 0.00, P <0.05 between groups. Conclusion The animal model of Clostridium perfringens with different concentrations of Clostridium perfringens at different stages of hind limb gas gangrene infection in mice was successfully established, which laid the foundation for understanding the pathogenesis and treatment of gas gangrene.