肿瘤研究常用动物小鼠病原菌16s rDNA检测方法的建立  

The establishment of the pathogen 16s rDNA detection method in mice commonly used animal for cancer research

作　　者：时伟红[1] 魏政立[1] 郭晓冲[1] 刘佳[1] 吴腾飞[1] 郑志红[1] 

机构地区：[1]中国医科大学实验动物部,辽宁沈阳110001

出　　处：《现代肿瘤医学》2015年第8期1061-1064,共4页Journal of Modern Oncology

基　　金：辽宁省科学技术计划项目(编号:2013408001)

摘　　要：目的:建立肿瘤研究常用动物小鼠病原菌快速检测方法。方法:提取经常规方法已被鉴定的小鼠病原菌绿脓杆菌、溶血性链球菌和大肠埃希氏菌DNA,利用PCR技术扩增病原菌16s rDNA全长,经纯化后直接测序。利用BLAST软件从Gen Bank数据库中搜索相关菌株的序列进行序列比对和同源性分析,确定病原菌的种属。结果:测序结果与数据库中搜索到的相关菌株的序列进行序列比对显示,3种病原菌测序序列分别与绿脓杆菌、溶血性链球菌和大肠埃希氏菌序列一致。结论:16s rDNA PCR方法可准确地检测肿瘤研究常用动物小鼠病原菌绿脓杆菌、溶血性链球菌和大肠埃希氏菌感染,并且缩短了检测时间。Objective：To establish the pathogen detection method in mice commonly used animal for cancer re- search. Methods ：The DNA of Escherichia coli, Pseudomonas aeruginosa and hemolytic streptococcus confirmed by conventional methods were extracted. The full length of 16s rDNA was amplified with a set of universal primers. The PCR products were purified and sequenced directly. The related sequences were obtained from the GenBank database using BLAST search software and the species of the pathogen were identified according to the sequence analysis and homology comparison of the 16s rDNA sequences. Results ：The 16s rDNA sequences of the three pathogen were 100% concordance with the known standard nueleotide sequences of Escherichia coli, Pseudomonas aeruginosa and hemolytic streptococcus. Conclusion：The PCR method based on 16s rDNA was reliable and could identify the pathogen rapidly and accuratly.

关 键 词：小鼠 病原菌 16s RDNA 多聚酶链反应 

分 类 号：R73-3[医药卫生—肿瘤]