宏基因组测序法在药品污染细菌鉴定与溯源中的应用研究  

Evaluation of the Application Value of Metagenomic Sequencing in Identification and Tracing of Bacteria Contaminating Drugs

作　　者：严卓彦 谭宇翔 林吉恒 夏瑛瑛 孙瑛 崔洁 黄朱梁 YAN Zhuoyan;TAN Yuxiang;LIN Jiheng;XIA Yingying;SUN Ying;CUI Jie;HUANG Zhuliang(National Ocean Food Quality Inspection Center,Zhoushan Institute for Food and Drug Control,Zhoushan 316012,China;CAS Key Laboratory of Quantitative Engineering Biology,Shenzhen Institute of Synthetic Biology,Shenzhen Institutes of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,China)

机构地区：[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 [2]中国科学院深圳先进技术研究院,深圳合成生物学创新研究院,中国科学院定量工程生物学重点实验室,广东深圳518055

出　　处：《中国药学杂志》2024年第11期1041-1046,共6页Chinese Pharmaceutical Journal

基　　金：浙江省药监局科技计划项目资助(2023010)。

摘　　要：目的:评价宏基因组测序法在药品污染细菌鉴定和溯源中的应用。方法利用宏基因组测序法和分离培养法同时对两批中成药及检测实验室环境取样中的细菌进行鉴定和分型。宏基因组测序结果通过MetaPhlan和StrainSifter软件进行菌株水平的分析。结果经宏基因测序发现,药品与环境中均有大量的蜡样芽孢杆菌污染,且药品B18与环境样本HJ2中含有相同的蜡样芽孢杆菌菌组。培养法从药品和环境中共分离得到的5株蜡样芽孢杆菌,通过蜡样芽孢杆菌特异性聚合酶链式反应(PCR)扩增基因gyrA和panC序列构建NJ进化树,以及多位点序列分型(MLST),结果均显示药品B18中的蜡样芽孢杆菌菌株与环境样本HJ2的菌株同组。结论宏基因组测序技术是可靠的,且相比培养法,能够更全面发现具有潜在风险的细菌物种,同时也能准确地进行菌株水平的溯源分析,在药品污染菌鉴定与溯源中具有重要的应用价值。OBJECTIVE To investigate metagenomic sequencing in the identification and tracing of bacteria contaminating drugs,and evaluate its application value.METHODS Both metagenomic sequencing and isolation culturing were employed to identify and type bacteria in two batches of Chinese patent medicines and environmental samples from the testing laboratory.The metagenomic sequencing results were analyzed at the strain level using MetaPhlan and StrainSifter software.RESULTS The analysis revealed a substantial contamination of Bacillus cereus-like organisms in both medicines and environmental samples,with the presence of the same B.cereus-like strain in medicine B18 and environmental sample HJ2.Through isolation culturing,five B.cereus-like strains were obtained from the medicines and environment.A phylogenetic tree was constructed based on the gyrA and panC gene sequences amplified by B.cereus-specific PCR,and multilocus sequence typing(MLST)results,all of which demonstrated that the B.cereus-like strain in medicine B18 belonged to the same group as one strain from the environment.CONCLUSION These findings indicate that metagenomic sequencing technology is reliable and has advantage over isolation culturing in comprehensively detecting potentially hazardous bacterial species and accurately performing strain-level traceability analysis.It holds significant application value in identification and tracing of pharmaceutical contamination.

关 键 词：宏基因组测序 蜡样芽孢杆菌特异性PCR 多位点序列分型 药品污染 细菌鉴定 细菌溯源 

分 类 号：R917[医药卫生—药物分析学]