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.
出处
《中国卫生检验杂志》
CAS
北大核心
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)
关键词
生理生化
PCR
肺炎克雷伯菌
特异性
鉴定
Physiology and biochemistry
PCR
Klebsiella pneumoniae
Specificity
