Rapid Sterility Test Method For Cellular Therapy Product

细胞治疗产品的快速无菌测试方法

• 批准号: 6675275
• 负责人: Patrick R Murray
• 金额: --
• 依托单位: CLINICAL CENTER
• 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/6675275
• 关键词: antiseptic sterilization; biological products; diagnostic tests; evaluation /testing; human tissue; microorganism growth; tissue /cell culture

Sterility testing is an essential part of in-process and release testing for cellular therapy products. The FDA specifically recommends that sterility testing be performed as outlined in 21 CFR 610.12. Furthermore, because they are concerned that antibiotics may interfere with the accurate assessment of sterility testing, the FDA requires preliminary bacteriostasis and fungistasis testing according to the USP "<71> Sterility Test" on all samples containing antibiotics, including cells grown in antibiotic-containing media. The methods for sterility testing described in the CFR and USP standards were developed more than 25 years ago, and are labor-intensive and require incubation for 14 days. Since the time these methods were published, more sensitive and rapid methods have been developed for detection of microbial growth in various body fluids. These modern methods include automated systems for the detection of microbial growth in blood and other normally sterile fluids. Despite the fact many laboratories use these automated systems to assess sterility of cellular therapy products, the FDA has not sanctioned this application because there are no published data from any formal comparison of these newer methods with the CFR and USP methods. For these reasons, we have developed a validation protocol comparing the CFR and USP methods with two automated culture methods: the bioMerieux BacT/Alert system and the Becton Dickinson Bactec system. Cell products will be seeded with selected bacteria and fungi and then tested with each method. The test sensitivity and time to detect a positive culture will be assessed. The validation protocol has been submitted to the FDA for their comments and we anticipate this study will be initiated in the Fall, 2002. As supporting data for this FDA submission, we have initiated a preliminary comparison of the CFR and USP methods with the automated bioMerieux BacT/Alert system (currently used in the NIH Microbiology Laboratory). All cellular therapy products (N=187) that have been prepared during August-September were tested in both systems. A total of 7 products had growth detected in one or both systems: 3 products in the BacT/Alert system only, 2 products in the CFR/USP method only, and 2 products in both systems. The clinical significance of these positive cultures will be assessed. This preliminary study will also be used to define (1) the time to detection of a positive culture in the CFR/USP and BacT/Alert methods; and (2) the incidence of false-positive culture signals (e.g., the system appears positive [cloudy; positive signal by the automated system] but no organisms are recovered in the system).

无菌检查是细胞治疗产品过程中和放行检查的重要组成部分。FDA特别建议按照21 CFR 610.12所述进行无菌试验。此外，由于他们担心抗生素可能干扰无菌试验的准确评估，FDA要求根据USP“<71>无菌试验”对所有含抗生素的样品（包括在含抗生素培养基中生长的细胞）进行初步抑菌和抑真菌试验。CFR和USP标准中描述的无菌试验方法是在25年前开发的，并且是劳动密集型的，需要孵育14天。自从这些方法发表以来，已经开发出更灵敏和快速的方法来检测各种体液中的微生物生长。这些现代方法包括用于检测血液和其他正常无菌液体中微生物生长的自动化系统。尽管许多实验室使用这些自动化系统来评估细胞治疗产品的无菌性，但FDA尚未批准此应用，因为这些新方法与CFR和USP方法的任何正式比较都没有发表数据。出于这些原因，我们制定了一项验证方案，将CFR和USP方法与两种自动培养方法（bioMerieux BacT/Alert系统和Becton Dickinson Bactec系统）进行比较。细胞产品将接种选定的细菌和真菌，然后采用每种方法进行检测。将评估检测灵敏度和检测阳性培养物的时间。验证方案已提交给FDA征求意见，我们预计本研究将于2002年秋季启动。作为此次FDA提交的支持性数据，我们已开始将CFR和USP方法与自动化bioMerieux BacT/Alert系统（目前在NIH微生物实验室使用）进行初步比较。在两个系统中测试了8月至9月期间制备的所有细胞治疗产品（N=187）。在一个或两个系统中共检测到7种产品有生长：仅BacT/Alert系统中有3种产品，仅CFR/USP方法中有2种产品，两个系统中有2种产品。将评估这些阳性培养物的临床意义。该初步研究还将用于定义（1）CFR/USP和BacT/Alert方法中检测阳性培养物的时间;和（2）假阳性培养信号的发生率（例如，系统显示阳性[浑浊;自动化系统显示阳性信号]，但系统中未回收到微生物）。