摘要
目的为解决内源性感染细菌体内示踪问题，本实验采用ＰＵＣ１９质粒载体示踪加限制性内切酶酶切图谱分析法和荧光标记示踪法，对肠道移位细菌进行了动态观察。并对两种方法进行了比较。方法依据ＰＵＣ１９质粒特点设计了动物模型，１１０只Ｗｉｓａｒ大鼠致成３０％体表面积Ⅲ°烫伤，于６、１２、２４、４８ｈ和１２天分别活杀，做荧光显微镜检查和细菌培养，质粒提取和酶切。结果烧伤后肠道细菌可移位至淋巴结和肝脏，荧光标记方法细菌到达淋巴结（９５％）和肝脏（５７．５％）的百分率略高于ＰＵＣ１９示踪方法（７８．４％、５２．４％）。结论用ＰＵＣ１９质粒载体做示踪物可减少非特异性因素。
OBJECTIVE In order to solve problem of bacterial tracing in vivo, The PUC19 plasmid vector trace with restriction map analysis and fluorescence labelling bacteria methods were applied to study gut origin bacterial translocation. Two methods were compared at same time. METHOD According to the characteristic of PUC19 plasmid, a special animal model was desingned. 110 wistar rats received 30% TBSA fall thickness burns. On hours 6、12、24、48 and day 12 postburn ,injuried animal were killed. Tissue homogenates of liver and mesenteric lymph nodes were examined by fluorescence microscope, bacterial culture and solation of plasmid and restriction map analysis. RESULT Gut bacteria can translocated to mesenteric lymph node and liver. The percentage that bacteria reach mesenteric lymph node and liver of fluorescence labelling method were higher than PUC19 trace method. CONCLUSION The PUC19 plasmid trace method can reduce nonspecificity factors. It is a ideal method for studing mechanism of endogenous infection.
出处
《中华医院感染学杂志》
CAS
CSCD
1998年第3期145-147,共3页
Chinese Journal of Nosocomiology
基金
国家自然科学基金
