PCR结合生理生化技术鉴定天然植物化妆品中肺炎克雷伯菌实验研究  

Study on identification of Klebsiella pneumoniae in natural plant-based cosmetics by physiological and biochemical technology and PCR

作　　者：马雪艺 黄玲[2] 史茜[2] 伊力亚 朱漪 刘思聪[2] 

机构地区：[1]新疆农业大学食品与药品科学学院,新疆乌鲁木齐830052 [2]新疆出入境检验检疫局技术中心,新疆乌鲁木齐830063

出　　处：《中国卫生检验杂志》2014年第7期913-916,共4页Chinese Journal of Health Laboratory Technology

基　　金：国家质检总局课题项目(2012IK194)

摘　　要：目的建立新疆本地特色天然植物性化妆品中肺炎克雷伯菌检测分析方法。填补天然植物性原料化妆品中特有致病菌检测技术空缺。方法通过PCR结合生理生化技术方法进行研究。实验初步分离出天然植物性染发粉中致病微生物,对分离到的菌落进行纯化、形态观察和生理生化鉴定,得到肺炎克雷伯菌生化反应结果。针对肺炎克雷伯菌phoE段基因设计特异性引物,提取最终纯化的细菌纯培养物菌株DNA,经PCR技术扩增得到348 bp目的扩增产物。结果通过验证引物特异性结果,确认得到该菌属肺炎克雷伯菌结果。实验条件下,检测得到的基因序列同源性与肺炎克雷伯菌高达99%。结论该检测方法可以在48 h内完成对样品中肺炎克雷伯菌的检测,相比单一微生物理化检测方法,该检测体系效率高,精确度高。Objective To establish an analytical method for Klebsiella pneumoniae detection in Xinjiang local natural plant-based cosmetics,and fill the technology gap of specific pathogenic bacteria detection in natural plant ingredients in cosmetics. Methods The pathogenic microorganism were isolated from natural plant-based hair coloring powder for purification,morphological observation,physiological and biochemical identification. Specific primers of the Klebsiella pneumoniae phoE gene were designed for DNA extraction of the final purified bacterial pure culture and PCR amplification. Results The verification results of primer specificity showed that the pathogeic bacteria was Klebsiella pneumoniae. The gene sequence was homologous with Klebsiella pneumoniae as high as 99%. Conclusion This method can complete the detection of Klebsiella pneumoniae in samples in 48 h. Compared with the single microbial detection method and chemical detection method,this detection system have high efficiency and high precision.

关 键 词：生理生化 PCR 肺炎克雷伯菌 特异性 鉴定 

分 类 号：R378.996[医药卫生—病原生物学]