宏基因组测序法对林可霉素利多卡因凝胶中污染细菌的鉴定与溯源研究*

doi:10.16155/j.0254-1793.2024-0072

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药物分析杂志 ›› 2024, Vol. 44 ›› Issue (8) : 1407-1414. DOI: 10.16155/j.0254-1793.2024-0072

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宏基因组测序法对林可霉素利多卡因凝胶中污染细菌的鉴定与溯源研究^*

严卓彦¹, 谭宇翔², 夏瑛瑛¹

作者信息 +

Using metagenomic sequencing to identify and trace drug-contaminating bacteria in lincomycin hydrochloride and lidocaine hydrochloride gel^*

YAN Zhuo-yan¹, TAN Yu-xiang², XIA Ying-ying¹

Author information +

文章历史 +

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: 分别通过培养法、扩增子测序法和宏基因组测序法,对2批微生物限度控制菌检查中检出未知微生物的林可霉素利多卡因凝胶药剂进行菌种鉴定和溯源。培养法分离得到的细菌,通过革兰氏染色、生化反应、生物质谱法、16srDNA测序及芽孢杆菌特异性PCR方法进行菌种鉴定,并通过多位点序列分型(MLST)法对药品和环境中均检出的蜡样芽孢杆菌进行菌株分型。宏基因组测序结果通过MetaPhlan软件进行菌株水平的分析。结果: MLST与MetaPhlan结果均显示药品与检测环境中存在同一组蜡样芽孢杆菌,显示药品中的蜡样芽孢杆菌极有可能来自检测环境的污染。培养法与宏基因组测序法均在药品中检出污染微生物土壤短芽孢杆菌、类芽孢杆菌及新芽孢杆菌,但扩增子测序还在环境样本中检出上述细菌,因此不能排除药品中此类污染菌来自检测环境的可能性。结论: 宏基因组测序技术相比培养法,能够更全面发现具有潜在风险的微生物,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。

Abstract

Objective: To evaluate the application of metagenomic sequencing in identification and tracking of drug-contaminating bacteria. Methods: Both metagenomic sequencing and isolation culturing method were employed to identify and genotype bacteria in two batches of possibly contaminated gel medicines as well as the environmental samples from testing laboratory. Bacteria isolated by culturing method were identified through Gram's stain, biochemical reaction, MALIDI-TOF-MS, 16srDNA sequencing and Bacillus specific PCR. The detected Bacillus cereus were typed by multilocus sequence typing (MLST) .The metagenomic sequencing results were analyzed at the strain level using MetaPhlan software. Results: Both MLST and MetaPhlan analysis revealed a substantial contamination of Bacillus cereus in gel medicines as well as environmental samples, with the presence of the same Bacillus cereus group. Both metagenomic sequencing and culturing methods detected Paenibacillus, Brevibacillus and Neobacillus in gel medicines, but amplicon sequencing also detected these bacteria in environment which might be the source of contamination. Conclusion: These findings indicate that metagenomic sequencing technology is reliable and has the advantage over isolation culturing in comprehensively detecting potentially hazardous bacterial species and accurately performing strain-level traceability analysis. It holds significant application value in pharmaceutical contamination identification and traceability.

导出引用

严卓彦, 谭宇翔, 夏瑛瑛. 宏基因组测序法对林可霉素利多卡因凝胶中污染细菌的鉴定与溯源研究^*[J]. 药物分析杂志, 2024, 44(8): 1407-1414 https://doi.org/10.16155/j.0254-1793.2024-0072

YAN Zhuo-yan, TAN Yu-xiang, XIA Ying-ying. Using metagenomic sequencing to identify and trace drug-contaminating bacteria in lincomycin hydrochloride and lidocaine hydrochloride gel^*[J]. Chinese Journal of Pharmaceutical Analysis, 2024, 44(8): 1407-1414 https://doi.org/10.16155/j.0254-1793.2024-0072

中图分类号： R 917

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