16S rRNA基因文库方法在感染标本菌群分析中的应用  

Application of 16S rRNA Gene Library Method in Bacterium Flora Analysis of Infected Wound

作　　者：常玉梅[1] 刘海燕[1] 

机构地区：[1]解放军252医院中心实验室,河北保定071000

出　　处：《解放军医药杂志》2013年第1期57-59,共3页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

摘　　要：目的探讨16S rRNA基因文库方法对感染标本菌群的鉴定效果。方法采集70例创伤患者的感染伤口脓液或渗出液标本,使用通用引物扩增标本中所有细菌的16S rRNA基因片段,构建16S rRNA基因文库,挑取阳性克隆测序,分析感染细菌的数量与种类。结果从150个克隆子中检测7种不同酶切类型的克隆,鉴定出7类微生物,其中屎肠球菌种类最多,占88%,坚强肠球菌比例占2%,另5类序列与非可培养细菌类群的16S rRNA基因序列相似性较高,均占2%。结论感染标本中屎肠球菌含量丰富,应用16S rRNA基因文库的方法可有效分离到感染标本中的非可培养菌。Objective To investigate the identification effect of 16S rRNA gene library method in bacterium flora analysis of infected samples. Methods Fresh pus samples or exudate of 70 trauma patients were collected. The 16S rRNA gene cloning library was established with 16S rRNA gene fragment in amplifiable samples of bacterial universal primers. Positive clones were sequenced, and amount and category of infected bacterials were analyzed. Results The total of 150 clones were selected nand 7 restriction fragment length polymorphism （RFLP） types and 7 kind of microbe which were used for sequencing. Enterococcus faecium was the most one （88% of total clones） , Enterococcus （2% of total clones） , 5 of these sequences were closely related to the uncultured bacteria fungi of 16S rRNA gene sequence simi- larity （2% of total clones respectively）. Conclusion The resuhs indicate that the rich Enterococcus faecium, and 16S rRNA gene library method may be an effective method for isolating uncultured bacteria in trauma infected samples.

关 键 词：创伤和损伤 感染 基因表达调控 细菌 基因克隆文库方法 

分 类 号：R446.51[医药卫生—诊断学] R349.83[医药卫生—临床医学]