高风险注射剂生产企业制药用水系统微生物菌库的建立和分析

Establishment and Analysis of Microbial Library in Pharmaceutical Water System of High-risk Injection Manufacturer

关键词:  无菌药品生产企业  制药用水系统  微生物菌库  注射用水  纯化水

Key words:  sterile pharmaceutical manufacturer  pharmaceutical water system  microbial library  water for injection  purified water

摘要:

目的 建立某高风险注射剂生产企业制药用水系统微生物数据库，为企业制药用水系统污染微生物的有效控制和溯源调查提供指导。方法 连续4个季度对某大容量注射剂生产企业的纯化水系统和注射用水系统监控收集微生物，采用基于16S rRNA和ITS rDNA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，建立企业制药用水系统的微生物菌库。结果 共分离鉴定1 499株细菌和20株真菌，注射用水系统和纯化水系统的微生物分别占比9.0%和91.0%。纯化水系统中鞘氨醇单胞菌和甲基杆菌最多，分别占全部分离菌的28.9%和25.2%。注射用水系统中葡萄球菌和微球菌较多，分别占全部收集菌的24.9%和18.2%。纯化水系统中污染的微生物主要为革兰氏阴性菌，占污染微生物总数的67.0%。注射用水系统中污染的微生物主要为革兰氏阳性菌，占污染微生物总数的60.3%。制药用水系统中较为常见的真菌为杂色曲霉菌和产黄青霉。此外，经分析发现，纯化水系统的微生物在第二季度和第三季度数量占比较多，占全年微生物总量67.8%，第一季度和第四季度微生物数量则下降；注射用水系统微生物数量则在第二季度较多，其余3个季节数量差不多。结论 建立制药用水系统微生物菌库可指导制药用水系统的运行维护及消毒灭菌，同时也为高风险药品生产企业的微生物污染事件提供溯源调查基础。

Abstract:

OBJECTIVE To establish a microbiological database for a pharmaceutical water system in the high-risk injection manufacturer, so as to provide guidance for effective control and traceability investigation of microorganisms polluted by the pharmaceutical water system. METHODS Microorganisms were monitored and collected in the purified water system and water for injection system of large-capacity injection manufacturer for four consecutive quarters, 16S rRNA and ITS rDNA sequencing methods were used to identify contaminated microorganisms. Statistical analysis was performed by combining the information of microbial species and source. A microorganisms library of pharmaceutical water system was established. RESULTS It was shown that 1 499 strains of bacteria and 20 strains of mold were isolated from pharmaceutical water system. Microorganisms in the water for injection and purified water systems accounted for 9.0% and 91.0%, respectively. Sphingomonas and Methylobacteria were the most in purified water system, accounting for 28.9% and 25.2%, respectively. Staphylococcus and Micrococcus were most in the water for injection system, accounting for 24.9% and 18.2%, respectively. The contaminated microorganisms in the purified water system were mainly Gram-negative bacteria, accounting for 67.0%. The contaminated microorganisms in the water for injection system were mainly Gram-positive bacteria, accounting for 60.3%. The more common fungi in pharmaceutical water system were Aspergillus versicolor and Penicillium chrysogenum. In addition, the analysis found that the number of microorganisms in the purified water system accounted for more in the second and third quarters, accounting for 67.8% of the total microorganisms in the year, and the number of microorganisms in the first and fourth quarters decreased. The number of microorganisms in the water for injection system was higher in the second quarter, and the numbers were similar in the remaining three seasons. CONCLUSION Establishing a microbial library for a pharmaceutical water system can guide the operation and maintenance of pharmaceutical water system and disinfection, and also can provide a source of traceability investigation for microbial contamination incidents in high-risk pharmaceutical manufacturer.