特异性PCR及MLST法对中成药常见污染芽孢杆菌的鉴定及溯源研究
Identify and Trace Contaminated Bacteria in Chinese Patent Drugs by Specific PCR and MLST Methods
严卓彦1
作者信息
- 1. 舟山市食品药品检验检测研究院 国家海洋食品质量检验检测中心,浙江 舟山 316012
- 折叠
摘要
目的 研究中成药中常见污染细菌的分离鉴定及溯源分析方法.方法 结合微生物限度检查工作的实例,对检出污染菌的34批中成药及检测环境中分离得到的细菌进行鉴定,通过生物质谱法初筛、16S rDNA测序和芽孢杆菌特异性PCR法进行验证.对药品和环境中都检出的枯草芽孢杆菌和蜡样芽孢杆菌通过多位点序列分型(MLST)法进行溯源分析.结果 生物质谱法能够鉴定绝大多数污染细菌,但无法鉴别蜡样芽孢杆菌群及某些近缘枯草芽孢杆菌群.16S rDNA测序结合芽孢杆菌特异性PCR方法能够快速鉴定所有的污染细菌.MLST结果显示药品与检测环境中存在同一类型的蜡样芽孢杆菌及不同类型的枯草芽孢杆菌,提示药品中检出的蜡样芽孢杆菌可能来源于检测环境.结论 中成药中绝大多数污染细菌为芽孢杆菌,芽孢杆菌特异性PCR及MLST法可以作为药品中污染芽孢杆菌的快速鉴定和溯源方法.
Abstract
Objective To study methods for identification and tracing contaminating bacteria in Chinese patent drugs.Methods In the daily work of microbial enumeration tests,bacteria cultured from 34 batches of Chinese patent drugs were identifed through MALDI-TOF-MS,16S rDNA sequencing and Bacillus specific PCR method.MLST methods were used to type Bacillus subtilis and Bacillus cereus detected both in drugs and lab environment.Results MALID-TOF-MS could identify most bacteria but not Bacillus cereus group and some Bacillus substilis groups which could be discriminated by 16S rDNA sequencing and Bacillus specific PCR method.MLST results showed there were same groups of Bacillus cereus in drugs and environment but different groups of Bacillus subtilis between them.Conclusion Most of the contaminating bacteria in Chinese patent drugs are Bacillus genus.Thus Bacillus specific PCR and MLST methods are quick and efficient ways to identify and trace contaminating bacteria in Chinese patent drugs.
关键词
芽孢杆菌特异性PCR/多位点序列分型/16SrDNA测序法/生物质谱法Key words
Bacillus specific PCR/MLST/16S rDNA sequencing/MALDI-TOF-MS引用本文复制引用
出版年
2024
NETL
NSTL
万方数据