稳定生物发光金葡菌临床株假体周围关节感染动物模型的构建

发布时间：2018-05-08 22:30

本文选题：金黄色葡萄球菌 + 感染　；参考：《中华关节外科杂志(电子版)》2016年04期

【摘要】：目的建立稳定型生物发光临床株金黄色葡萄球菌假体周围关节感染(PJI)动物模型并探讨其可行性及应用价值。方法应用噬菌体Phi11侵染带Lux发光基因的标准菌株BD1652,并通过噬菌体转导将Lux基因导入PJI临床菌株金葡菌ST1792基因组DNA,进而构建成稳定型生物发光临床菌株ST1792-Lux。通过抽纸牌,将10只健康BALB/c小鼠随机均分为实验组和对照组,使用克氏针作为两组模型动物的左膝关节假体。术中实验组左膝关节内注入10μl(105CFU)ST1792-Lux菌液,对照组注入等量0.9%氯化钠溶液,术后1、3、7、14 d,采用活体成像系统(IVIS)检测生物发光强度随时间变化;并于术后2周观察小鼠左膝关节病理学变化。以非配对t检验对ST1792-Lux和ST1792的生物膜形成能力和同一时间点实验组与对照组小鼠左膝平均生物发光强度进行比较分析。结果经临床株筛选,噬菌体Phi11转导获得生物发光菌株"ST1792-Lux",无抗生素筛选条件下连续传代培养48 h,ST1792-Lux稳定发光;Lux基因重组于ST1792基因组DNA内,较之野生株ST1792,ST1792-Lux在生长能力和不同培养条件下成膜能力(tTSB=1.16,1.29;tTSBG=0.38,0.31;P均0.05)方面差异无统计学意义。IVIS检测示,术后每个时间点(1、3、7和14 d),实验组较对照组的左膝平均生物发光强度均明显增高(t=9.13,10.72,14.48,7.46;P均0.05)。结论本研究成功建立了稳定生物发光临床株S.aureus PJI动物模型,其稳定性高、重复性好,可用于临床株S.aureus PJI的致病分子机制和各种干预措施有效性的体内研究。
[Abstract]:Objective to establish a stable bioluminescence clinical strain of Staphylococcus aureus periprosthetic joint infection (PJI) animal model and explore its feasibility and application value. Methods the standard strain BD1652 with Lux luminescence gene was infected with phage Phi11, and the Lux gene was transfected into ST1792 genomic DNA of PJI clinical strain ST1792 by phage transduction, and then the stable bioluminescence clinical strain ST1792-Lux. was constructed. By playing cards, 10 healthy BALB/c mice were randomly divided into experimental group and control group. Kirschner needle was used as the prosthesis of left knee joint in two groups of model animals. 10 渭 l(105CFU)ST1792-Lux bacterial solution was injected into the left knee joint of the experimental group and 0.9% sodium chloride solution was injected into the control group. The bioluminescence intensity was measured by living imaging system at 14 days after the operation, and the pathological changes of the left knee joint were observed at 2 weeks after operation. The biofilm forming ability of ST1792-Lux and ST1792 and the average bioluminescence intensity of the left knee of the experimental group and the control group at the same time point were compared and analyzed by non-paired t test. Results the bioluminescent strain "ST1792-Lux" was obtained by phage Phi11 transduction. After 48 h culture without antibiotic screening, ST1792-Lux gene was recombined into ST1792 genomic DNA. Compared with wild strain ST1792, ST1792-Lux had no significant difference in growth ability and membrane forming ability under different culture conditions. The mean bioluminescence intensity of the left knee in the experimental group was significantly higher than that in the control group on the 7th and 14th day after operation. The mean bioluminescence intensity of the left knee in the experimental group was significantly higher than that in the control group. Conclusion the animal model of stable bioluminescence clinical strain S.aureus PJI has been successfully established in this study. The animal model is stable and reproducible. It can be used to study the pathogenetic molecular mechanism of clinical strain S.aureus PJI and the effectiveness of various intervention measures in vivo.
【作者单位】：上海交通大学附属第六人民医院骨科;上海交通大学附属第六人民医院检验科;
【基金】：国家自然科学基金(81472108)
【分类号】：R-332;R687.4;R318.17

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