title: Sterility Testing of Stem Cell Products by Broad-Range Bacterial 16S Ribosomal DNA Polymerase Chain Reaction.
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.
authors.affiliation: Blood Transfusion Division, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan otokuno@cira.kyoto-u.ac.jp.
authors.forename: Osamu
authors.initials: O
authors.lastname: Tokuno
authors.affiliation: Department of Pediatrics, and.
authors.forename: Akira
authors.initials: A
authors.lastname: Hayakawa
authors.affiliation: Department of Pediatrics, and.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yanai
authors.affiliation: Department of Pediatrics, and.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Mori
authors.affiliation: Clinical Laboratory, Kobe University Hospital, Kobe, Japan, and.
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Ohnuma
authors.affiliation: Blood Transfusion Division.
authors.forename: Ayumi
authors.initials: A
authors.lastname: Tani
authors.affiliation: Blood Transfusion Division.
authors.forename: Hironobu
authors.initials: H
authors.lastname: Minami
authors.affiliation: Blood Transfusion Division.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Sugimoto
mesh_terms: D004269:DNA, Bacterial / Q000032:analysis*; D006801:Humans; D007246:Infertility; D012333:RNA, Messenger / Q000378:metabolism; D012336:RNA, Ribosomal, 16S / Q000378:metabolism*; D013234:Stem Cells / Q000378:metabolism*
chemical_list: D004269:DNA, Bacterial; D012333:RNA, Messenger; D012336:RNA, Ribosomal, 16S
keywords: 16S rDNA; BacT/ALERT; bacterial contamination; blood products; stem cells