无菌药品生产企业过程污染微生物菌株库的建立及数据分析

目的：建立某无菌药品生产企业过程污染微生物菌株库，为无菌药品生产过程污染微生物有效控制和溯源调查提供指导。方法：对原辅料、包装材料、生产人员、生产环境、制药用水和检验实验室等环节建立污染微生物监控方案，采用基于16S rRNA序列比对等方法鉴定污染微生物，结合微生物种属和来源信息进行分析，建立覆盖药品生产过程的污染微生物菌株库。结果：共分离鉴定292株微生物，其中生产环境污染率最高，占全部污染微生物来源的72.6%；其次是生产用水（13.4%）、原辅料（10.3%）、生产人员（3.77%）。值得关注的是，A级洁净生产区检测到30株微生物。污染微生物主要分布于44个属、95个种，表明药品生产过程污染微生物种类复杂。其中葡萄球菌污染率最高，占全部污染微生物的40.25%，其次是微球菌（11.20%）、芽孢杆菌（8.30%）和不动杆菌（5.81%）。此外，经分析发现，部分污染微生物分布具有空间特异性，如芽孢杆菌主要存在于原辅料活性炭中，而不动杆菌则主要存在于注射用水中。结论：无菌药品生产过程中多个环节都存在微生物污染，且污染微生物种类复杂，建立覆盖整个生产过程的微生物监控和污染微生物数据库，对药品微生物质量控制和溯源调查分析具有非常重要的作用。

Objective: To establish a contaminated microorganisms library for sterile pharmaceutical manufacturing processes, so as to provide guidance for effective microbial control and traceability of contaminated microorganisms. Methods: Microbiological monitoring was conducted for raw materials and products, packaging materials, production personnel, production environment, pharmaceutical water, and microbiology laboratory. 16S rRNA sequencing method was used to identify the contaminating microorganisms. Statistical analysis was performed by combining the information of microbial species and source. A contaminated microorganisms library covering the pharmaceutical production process was established. Results: It was shown that 292 strains were isolated from pharmaceutical manufacturing process. Among them, the production environment showed the highest microbial positive rate, accounting for 72.6%, followed by production water 13.4%, raw materials 10.3% and production personnel 3.77%. Of concern, 30 strains were detected in the Class A clean area. All contaminated microorganisms in this study were distributed in 44 genera and 95 species, demonstrating that the contaminated microorganisms in the pharmaceutical production process are abundant. Staphylococcus (40.25%) and Micrococcus (11.20%) were the dominated microorganisms, followed by Bacillus (8.3%) and Acinetobacter (5.8%). By further analysis, it was found that the distribution of some contaminated microorganisms was spatially specific. For example, Bacillus strains were mainly found in carbon materials, while Acinetobacter strains were mainly present in injection water. Conclusion: Microbial contamination can be found in multiple areas of drug manufacturing process. Therefore, mapping the distribution of contaminated microorganisms in the drug manufacturing process has a very important role in the drugs quality control and microbiological traceability investigation.

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