Sterility Testing of Stem Cell Products by Broad-Range Bacterial 16S Ribosomal DNA Polymerase Chain Reaction
| Autor: | Akira Hayakawa, Takeshi Mori, Kenichiro Ohnuma, Tomoko Yanai, Ayumi Tani, Takeshi Sugimoto, Hironobu Minami, Osamu Tokuno |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
DNA
Bacterial medicine.medical_specialty Microbiological culture Sterility Stem Cells Biochemistry (medical) Clinical Biochemistry Transfusion medicine Biology 16S ribosomal RNA Molecular biology Microbiology law.invention law Infertility RNA Ribosomal 16S medicine Humans RNA Messenger Stem cell Ribosomal DNA Polymerase chain reaction Whole blood |
| Zdroj: | Laboratory Medicine. 46:34-41 |
| ISSN: | 1943-7730 0007-5027 |
| DOI: | 10.1309/lmkt4p9ffi2bbsiu |
| Popis: | 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. * rDNA : ribosomal DNA PBSC : peripheral blood stem cell TSB : Trypticase Soy Broth SCD : Soybean-Casein Digest CFU : colonyforming units NCs : nonspiked controls UV : ultraviolet G-CSF : granulocyte–colony stimulating factor FDA : United States Food and Drug Administration |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|