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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.

journal_name

Lab Med

journal_title

Laboratory medicine

authors

Tokuno O,Hayakawa A,Yanai T,Mori T,Ohnuma K,Tani A,Minami H,Sugimoto T

doi

10.1309/LMKT4P9FFI2BBSIU

subject

Has Abstract

pub_date

2015-01-01 00:00:00

pages

34-41

issue

1

eissn

0007-5027

issn

1943-7730

pii

46/1/34

journal_volume

46

pub_type

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