Resumo

Introduction and Objectives: Nowadays, the harmonized sterility test method described on pharmacopoeias requires an incubation period of 14 days. During this period the product stays on quarantine waiting the result for release, which demands warehousing and inventory costs. Rapid Microbiological Methods (RMM`s)have been accepted around the world only after a complete validation, which requires results equivalent or better compared to those obtained by compendial method. The RMM based on carbon dioxide detection has as principle the detection of pad color change due to carbon dioxide release by microorganism metabolism. Regarding regulatory perspectives, there is a great resistance to accept non compendial methods, once it is a so critical issue. Given the above, this work has the objective of evaluating comparatively the time of detection of three different products intentionally contaminated with low levels of challenge microorganisms, in order to prove the method is equivalent or superior than compendial methods and to contribute to greater regulatory acceptance for application in high volume parenteral solution sterility testing. Material and Method: BacT/Alert 3D system was used as RMM. Products tested: Buffer saline solution, Metronidazol solution and Polieletrolitic dialysis concentrate solution. Challenge Microrganisms: Staphylococcus aureus (ATCC 6538), Bacilus subtilis (ATCC 6633), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 8739 ), Clostridium sporogenes (ATCC 19404), Candida albicans (ATCC 10231) and Aspergillus brasiliensis (ATCC 16404). Inoculum concentration: 20CFU/sample; 2 CFU/sample; 0,2CFU/sample. Incubation temperature: 32,5 ± 0,5 °C. Period of evaluation of the tests: 14 days. Aseptic samples of the three products were intentionally contaminated with BioBalls® SingleShot of challenge microorganisms, according to inoculum concentration desired, filtered in a 45µm pore membrane, inoculated into Soybean-Casein Digest Medium, and Fluid Tioglycollate Medium. After 18h aliquotes of 5mL were taken from inoculated culture media and inoculated into BacT/Alert 3D culture media bottles. Results: Results shows that the proportion of positive/false-negative samples among the different concentrations remained similar between the RMM and the compendial test, and the time of detection of the samples in RMM was statistically lower (p<0,05) than compendial method.