Analysis of the Microstar Rapid Sterility Test Method

Microstar快速无菌检测方法分析

• 批准号: 6546253
• 负责人: M C ANDERSON
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6546253
• 关键词: antisepsis; biological products; bioluminescence; biotechnology; evaluation /testing; germ free condition; method development; microorganism culture; microorganism growth; public health; tissue /cell culture

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.

摘要：对于要求无菌的生物制品，联邦法规规定的测试需要14天的潜伏期，并根据对接种产品的介质的目测检查进行观察，以合理确定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。微星系统由米利波尔公司制造，利用生物发光检测活的污染生物体，是根据实验室目前使用的现有米利波尔灭菌设备和用品选择进行评估的。今年的研究涉及将不到50 cfu的选定微生物注射到200毫升的目标生物制品中，然后对该产品进行传统的无菌测试和杂交无菌测试，其中每天使用Microstar方法对少量接种的介质进行检测。与传统的无菌检测相比，Microstar程序可以提前48-72小时检测到污染。在污染程度、生物体类型和检测时间之间的相关性方面，正在继续努力提高微星方法的灵敏度。今年已经尝试开始评估Microstar系统，用于检测基于细胞的产品中的污染物。这些产品必须在14天潜伏期结束前注入患者体内，这些产品可能会受到Microstar系统验证的有利影响。几种细胞类型的假制剂被少量的细菌CFU污染；然而，专门猝灭哺乳动物或动物细胞的生物发光的方法和试剂需要进一步研究。