经小鼠尾静脉注射诱导肺炎克雷伯菌血流感染模型的建立及评价

发布时间：2018-03-21 02:32

本文选题：肺炎克雷伯菌　切入点：毒力因子　出处：《中华医院感染学杂志》2017年06期 　论文类型：期刊论文

【摘要】：目的建立可靠的肺炎克雷伯菌血流感染模型,为细菌性血流感染早期诊断相关研究提供可靠的依据。方法选择肺炎克雷伯菌标准株ATCC 700603为实验菌株,将不同浓度的细菌悬液通过尾静脉注入小鼠体内,按Karber法计算LD50,以1/2LD50浓度注射小鼠,通过其体征变化、体重变化、血培养、分子生物学试验、H·E染色等指标综合评价模型,验证造模效果。结果小鼠LD50为1.11×109 CFU/ml,菌液感染小鼠1h后开始出现立毛,活动减少;感染前体重为(33.60±1.21)g,注射菌液后1天内体重下降至(28.86±1.42)g,跟对照组比较有明显差别(P0.05),7天后恢复正常水平(33.17±1.72)g;正常小鼠白细胞为(1.42±0.66)×109/L,注射菌液3h后升至(2.52±1.01)×109/L,6h升至(3.08±0.85)×109/L,跟对照组比较有明显差别(P0.05),2天后白细胞进一步升至(6.88±3.11)×109/L,7天后仍无明显下降;小鼠全血培养结果表明前2天任意时间血培养结果为阳性;毒力因子引物扩增疑似菌DNA,结果表明小鼠血流感染病原菌均为肺炎克雷伯菌;注射菌液1天后小鼠肺泡明显渗出、水肿、白细胞浸润;2天后肺结构逐渐恢复,肝恢复速度较肺慢。结论建立了小鼠肺炎克雷伯菌血流感染模型,能够为血流感染的早期诊断和不同病原菌感染的鉴别诊断等研究提供了可靠的动物模型。
[Abstract]:Objective to establish a reliable blood stream infection model of Klebsiella pneumoniae, and to provide reliable basis for the early diagnosis of bacterial blood flow infection. Methods ATCC 700603, a standard strain of Klebsiella pneumoniae, was selected as the experimental strain. Different concentrations of bacterial suspensions were injected into mice via tail vein, LD50 was calculated by Karber method, and 1 / 2 LD50 was injected into mice. A comprehensive evaluation model was established based on the changes of physical sign, body weight, blood culture, molecular biology test and H 路E staining. Results the LD50 of mice was 1.11 脳 109CFU / ml. The body weight before infection was 33.60 卤1.21g, and the body weight dropped to 28.86 卤1.42g within 1 day after injection of bacterial solution, which was significantly different from that of the control group, and returned to normal level after 7 days (33.17 卤1.72g). The leukocyte of normal mice was 1.42 卤0.66 脳 109 / L, and rose to 2.52 卤1.01 脳 109L / L and 3.08 卤0.85 脳 109L / L after 3 h injection of bacterial solution, and 3.08 卤0.85 脳 109L / L after 3 h injection of bacterial solution, compared with the control group, the white blood cells of normal mice were 1.42 卤0.66 脳 109 / L, respectively. After 2 days, the leukocyte level increased to 6.88 卤3.11) 脳 10 ~ 9 / L ~ (-1) and no significant decrease was found after 7 days. The results of whole blood culture of mice showed that the results of blood culture were positive at any time of the first 2 days; the DNA of suspected bacteria was amplified by the primers of virulence factor. The results showed that the pathogenic bacteria of blood stream infection in mice were Klebsiella pneumoniae. After 2 days of edema, leukocyte infiltration, the lung structure gradually recovered, and the recovery rate of liver was slower than that of lung. Conclusion the blood stream infection model of Klebsiella pneumoniae was established in mice. It can provide a reliable animal model for the early diagnosis of blood flow infection and the differential diagnosis of different pathogens.
【作者单位】：温州医科大学检验医学院生命科学学院;解放军总医院临床检验科;解放军总医院微生物科;
【基金】：国家重大科学仪器专项项目任务(2012YQ18011708)
【分类号】：R-332;R515

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