16S核糖体RNA基因片段分析及在药品葡萄球菌污染控制应用中的探讨  

A detailed analysis of 16S ribosomal RNA gene segments for identification of Staphylococcus contaminants in pharmaceutical quality control

作　　者：冯震[1] 李芳[1] 宋明辉 洪小栩[2] 许华玉[2] 刘浩[1] 杨美成[1] FENG Zhen;LI Fang;SONG Ming-hui;HONG Xiao-xu;XU Hua-yu;LIU Hao;YANG Mei-cheng(Shanghai Institute for Food and Drug Control, Shanghai 201203, China;Chinese Pharmacopoeia Commission, Beijing 100061, China)

机构地区：[1]上海市食品药品检验所,上海201203 [2]国家药典委员会,北京100061

出　　处：《药物分析杂志》2018年第4期680-687,共8页Chinese Journal of Pharmaceutical Analysis

基　　金：2015年国家药典委员会标准提高研究项目:<分子生物学技术用于药品质量控制标准核酸序列数据库的建立;分子生物学技术用于药品质量控制标准化技术指导原则的制定>;2016年上海市科委技术标准专项:<药品中微生物鉴定通用DNA条形码国家标准的建立>(16DZ0500202)

摘　　要：目的:建立细菌16S核糖体RNA基因片段分析的规范化方法,探讨其在药品葡萄球菌污染控制中的应用。方法:对Genbank数据库中用于常见细菌污染物鉴定的核酸序列进行筛选、定位和分区比较分析,建立具有鉴定和分类意义的标准核酸序列片段;以葡萄球菌为例进行菌株的核酸测序鉴定,分析菌株间的聚类关系和核酸序列的多样性。结果:16S rRNA基因中的V1~V3可变区具有显著的鉴定和分类意义,可实现葡萄球菌属20个"种"水平的鉴定。结论:本文对16S rRNA基因核酸测序鉴定规范化方法及在制药行业中葡萄球菌污染控制的应用进行探讨,为细菌核酸测序鉴定标准化和标准核酸序列数据库建设提供研究思路和参考依据。Objective：To analyze variable region segments of bacterial 16 S ribosomal RNA gene,and optimize criteria nucleotide sequence for Staphylococcus identification in drug production and examination environment.Methods：Nucleotide sequences of critical bacterial contaminants in drug production from Genbank nucleotide database were screened,positioned and analyzed,and criteria sequence was established.Staphylococcus isolates were collected and identified using sequencing method,and its sequence was analyzed by gene clustering method and polymeric method.Results：V1-V3 region sequences of 16 S ribosomal RNA gene had the perfect capability in bacterial identification,and this method could specifically classify 20 species of Staphylococcus.Conclusion：The criteria gene sequencing method for identifying bacterial contaminants were analyzed and discussed,it may provide a reference data in establishing criteria procedure and nucleotide sequence database for drug quality control.

关 键 词：16S RRNA基因 序列片段 标准核酸序列 葡萄球菌 污染控制 

分 类 号：R917[医药卫生—药物分析学]