Osamu Tokuno1, Akira Hayakawa2, Tomoko Yanai2, Takeshi Mori2, Kenichiro Ohnuma3, Ayumi Tani4, Hironobu Minami4, Takeshi Sugimoto4.
Abstract
OBJECTIVE: To evaluate broad-range 16S ribosomal DNA (rDNA) polymerase chain reaction (PCR) as a rapid screening tool to detect bacterial contamination of stem-cell products.
METHODS: We performed the evaluation using whole blood spiked with serially diluted bacterial-type strains. Detection sensitivity was defined as the bacterial concentration for which all replicates were positive at each concentration (100% detection). We tested the sterility of 29 bags of autologous peripheral blood stem cell (PBSC) products harvested at our facility using the 16S rDNA PCR method.
RESULTS: The detection sensitivity of 16S rDNA PCR in spiked whole blood was 10¹ to 10² colony-forming units (CFU) per mL, depending on the bacterial strain. We detected no amplified 16S rDNA among the PBSCs we used in this study. The BacT/ALERT automated bacterial culture system that we used also showed no positive signals in any of the PBSCs tested.
CONCLUSIONS: Our data indicate that bacterial 16S rDNA PCR is a useful alternative for rapid sterility testing, not only for blood products used in transfusion medicine but also for stem-cell products used in regenerative medicine. Copyright© by the American Society for Clinical Pathology (ASCP).
OBJECTIVE: To evaluate broad-range 16S ribosomal DNA (rDNA) polymerase chain reaction (PCR) as a rapid screening tool to detect bacterial contamination of stem-cell products.
METHODS: We performed the evaluation using whole blood spiked with serially diluted bacterial-type strains. Detection sensitivity was defined as the bacterial concentration for which all replicates were positive at each concentration (100% detection). We tested the sterility of 29 bags of autologous peripheral blood stem cell (PBSC) products harvested at our facility using the 16S rDNA PCR method.
RESULTS: The detection sensitivity of 16S rDNA PCR in spiked whole blood was 10¹ to 10² colony-forming units (CFU) per mL, depending on the bacterial strain. We detected no amplified 16S rDNA among the PBSCs we used in this study. The BacT/ALERT automated bacterial culture system that we used also showed no positive signals in any of the PBSCs tested.
CONCLUSIONS: Our data indicate that bacterial 16S rDNA PCR is a useful alternative for rapid sterility testing, not only for blood products used in transfusion medicine but also for stem-cell products used in regenerative medicine. Copyright© by the American Society for Clinical Pathology (ASCP).
Keywords:
16S rDNA; BacT/ALERT; bacterial contamination; blood products; stem cells
Mesh：
Substances：
Year: 2015
PMID： 25617390 DOI： 10.1309/LMKT4P9FFI2BBSIU
Source DB: PubMed Journal: Lab Med ISSN： 0007-5027