Analysis of the Microstar Rapid Sterility Test Method

摘要：对于要求无菌的生物制品，联邦法规规定的测试要求有14天的潜伏期，并根据对产品接种介质的目视检查得出结论，合理确信产品没有外来细菌或真菌污染。一组新出现的“快速”方法表明，用其他方法可以在不到14天的时间内检测到这种细菌污染。Millipore公司生产的Microstar系统利用生物发光来检测有活力的污染生物体，根据实验室目前对现有Millipore Steritest设备和用品的使用情况，选择了Microstar系统进行评估。今年的研究涉及将少于50 CFU的选定生物体注射到200毫升的目标生物制品中所引起的污染；然后对产品进行传统的无菌测试和混合无菌测试，其中每天使用MicroStar方法对少量接种培养基进行分析。与传统的无菌检测相比，MicroStar程序可提前48-72小时检测到污染。目前正在继续努力提高MicroStar方法在污染程度、生物体类型和检测时间之间的相关性方面的灵敏度。今年开始尝试评估MicroStar系统对细胞基产品中污染物的检测。这些产品必须在14天的潜伏期完成之前输注到患者体内，可能会受到MicroStar系统验证的有利影响。几种细胞类型的假制剂被低数量的细菌CFU污染；然而，特异性猝灭哺乳动物或动物细胞生物发光的方法和试剂需要进一步研究。

Summary: For biological products that are required to be sterile, testing mandated by the Code of Federal Regulations requires a 14 day incubation period, with observations based on visual examination of product-inoculated media, to conclude with reasonable confidence that the product is free from extraneous bacterial or fungal contamination. A group of newly emerging "rapid" methods suggests that such bacterial contamination can be detected in less than 14 days by other methods. The Microstar system, made by Millipore Corporation, utilizing bioluminescence to detect viable contaminating organisms, was selected for evaluation based on the laboratory's current use of existing Millipore Steritest equipment and supplies. Studies this year involved contamination introduced by injecting less than 50 CFU of selected organisms into a 200 ml vial of a target biological product; the product was then subjected to traditional sterility testing and a hybrid sterility test where small quantities of the inoculated media was assayed daily using the MicroStar methodology. The contamination was detected 48-72 hours earlier with the MicroStar procedure as compared to traditional sterility test. Work is continuing to increase the sensitivity of the MicroStar methodology, in terms correlation between contamination levels, types of organisms and detection time. Attempts were made this year to begin evaluating the MicroStar system for the detection of contaminants in cell based products. These products, that must be infused into patients prior to the completion of the 14 day incubation period, could be favorably impacted by the validation of the MicroStar System. Sham preparations of several cell types were contaminated with low numbers of bacterial CFU's; however, methods and reagents to specifically quench the bioluminescence of the mammalian or animal cells require further research.