Sterility Testing of Stem Cell Products by Broad-Range Bacterial 16S Ribosomal DNA Polymerase Chain Reaction
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 sensiti...
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| Veröffentlicht in: | Laboratory medicine 2015-02, Vol.46 (1), p.34-41 |
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| container_title | Laboratory medicine |
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| creator | Tokuno, Osamu Hayakawa, Akira Yanai, Tomoko Mori, Takeshi Ohnuma, Kenichiro Tani, Ayumi Minami, Hironobu Sugimoto, Takeshi |
| description | 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. |
| doi_str_mv | 10.1309/LMKT4P9FFI2BBSIU |
| format | Article |
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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.</description><identifier>ISSN: 0007-5027</identifier><identifier>EISSN: 1943-7730</identifier><identifier>DOI: 10.1309/LMKT4P9FFI2BBSIU</identifier><identifier>PMID: 25617390</identifier><language>eng</language><publisher>Oxford, UK: Oxford University Press</publisher><subject>Bacteria ; Deoxyribonucleic acid ; DNA ; DNA, Bacterial - analysis ; Gene amplification ; Humans ; Infertility ; Medicine ; Methods ; Process controls ; RNA, Messenger - metabolism ; RNA, Ribosomal, 16S - metabolism ; Stem cells ; Stem Cells - metabolism</subject><ispartof>Laboratory medicine, 2015-02, Vol.46 (1), p.34-41</ispartof><rights>Copyright© by the American Society for Clinical Pathology (ASCP) 2015</rights><rights>Copyright© by the American Society for Clinical Pathology (ASCP).</rights><rights>Copyright American Society for Clinical Pathology Winter 2015</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c405t-dfffd8f809f973e22aac42609221dadf3b0c7a1e631e6cc4ae6def4d4ecbc90b3</citedby><cites>FETCH-LOGICAL-c405t-dfffd8f809f973e22aac42609221dadf3b0c7a1e631e6cc4ae6def4d4ecbc90b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25617390$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tokuno, Osamu</creatorcontrib><creatorcontrib>Hayakawa, Akira</creatorcontrib><creatorcontrib>Yanai, Tomoko</creatorcontrib><creatorcontrib>Mori, Takeshi</creatorcontrib><creatorcontrib>Ohnuma, Kenichiro</creatorcontrib><creatorcontrib>Tani, Ayumi</creatorcontrib><creatorcontrib>Minami, Hironobu</creatorcontrib><creatorcontrib>Sugimoto, Takeshi</creatorcontrib><title>Sterility Testing of Stem Cell Products by Broad-Range Bacterial 16S Ribosomal DNA Polymerase Chain Reaction</title><title>Laboratory medicine</title><addtitle>Lab Med</addtitle><description>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.</description><subject>Bacteria</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Bacterial - analysis</subject><subject>Gene amplification</subject><subject>Humans</subject><subject>Infertility</subject><subject>Medicine</subject><subject>Methods</subject><subject>Process controls</subject><subject>RNA, Messenger - metabolism</subject><subject>RNA, Ribosomal, 16S - metabolism</subject><subject>Stem cells</subject><subject>Stem Cells - metabolism</subject><issn>0007-5027</issn><issn>1943-7730</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqFkctPwjAcxxujEUTvnkwTLyZm2hcbPQKKElEJj_PS9YEj24rtduC_twT0wMUmTfNtPt9vfg8ArjF6wBTxx8n724JN-Wg0JoPBfLw8AW3MGY2ShKJT0EYIJVEXkaQFLrxfB8l4TM5Bi3RjnFCO2qCY19rlRV5v4UL7Oq9W0BoYPks41EUBp86qRtYeZls4cFaoaCaqlYYDIXdGUUAcz-Esz6y3ZVBPH304tcW21E54DYdfIq_gTAc6t9UlODOi8Prq8HbAcvS8GL5Gk8-X8bA_iSRD3TpSxhjVMz3EDU-oJkQIyUiMOCFYCWVohmQisI5puFIyoWOlDVNMy0xylNEOuNvnbpz9bkJbaZl7GdoRlbaNT3HcJSyE0SSgt0fo2jauCtUFKhzOeUwDhfaUdNZ7p026cXkp3DbFKN1tIj3eRLDcHIKbrNTqz_A7-gDc7wHbbP6P-wE3DpLC</recordid><startdate>20150201</startdate><enddate>20150201</enddate><creator>Tokuno, Osamu</creator><creator>Hayakawa, Akira</creator><creator>Yanai, Tomoko</creator><creator>Mori, Takeshi</creator><creator>Ohnuma, Kenichiro</creator><creator>Tani, Ayumi</creator><creator>Minami, Hironobu</creator><creator>Sugimoto, Takeshi</creator><general>Oxford University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7RV</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8C1</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>KB0</scope><scope>M0S</scope><scope>M1P</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope></search><sort><creationdate>20150201</creationdate><title>Sterility Testing of Stem Cell Products by Broad-Range Bacterial 16S Ribosomal DNA Polymerase Chain Reaction</title><author>Tokuno, Osamu ; Hayakawa, Akira ; Yanai, Tomoko ; Mori, Takeshi ; Ohnuma, Kenichiro ; Tani, Ayumi ; Minami, Hironobu ; Sugimoto, Takeshi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c405t-dfffd8f809f973e22aac42609221dadf3b0c7a1e631e6cc4ae6def4d4ecbc90b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Bacteria</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA, Bacterial - analysis</topic><topic>Gene amplification</topic><topic>Humans</topic><topic>Infertility</topic><topic>Medicine</topic><topic>Methods</topic><topic>Process controls</topic><topic>RNA, Messenger - metabolism</topic><topic>RNA, Ribosomal, 16S - metabolism</topic><topic>Stem cells</topic><topic>Stem Cells - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Tokuno, Osamu</creatorcontrib><creatorcontrib>Hayakawa, Akira</creatorcontrib><creatorcontrib>Yanai, Tomoko</creatorcontrib><creatorcontrib>Mori, Takeshi</creatorcontrib><creatorcontrib>Ohnuma, Kenichiro</creatorcontrib><creatorcontrib>Tani, Ayumi</creatorcontrib><creatorcontrib>Minami, Hironobu</creatorcontrib><creatorcontrib>Sugimoto, Takeshi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Nursing & Allied Health Database</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Public Health Database</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Nursing & Allied Health Premium</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><jtitle>Laboratory medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tokuno, Osamu</au><au>Hayakawa, Akira</au><au>Yanai, Tomoko</au><au>Mori, Takeshi</au><au>Ohnuma, Kenichiro</au><au>Tani, Ayumi</au><au>Minami, Hironobu</au><au>Sugimoto, Takeshi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Sterility Testing of Stem Cell Products by Broad-Range Bacterial 16S Ribosomal DNA Polymerase Chain Reaction</atitle><jtitle>Laboratory medicine</jtitle><addtitle>Lab Med</addtitle><date>2015-02-01</date><risdate>2015</risdate><volume>46</volume><issue>1</issue><spage>34</spage><epage>41</epage><pages>34-41</pages><issn>0007-5027</issn><eissn>1943-7730</eissn><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.</abstract><cop>Oxford, UK</cop><pub>Oxford University Press</pub><pmid>25617390</pmid><doi>10.1309/LMKT4P9FFI2BBSIU</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Laboratory medicine, 2015-02, Vol.46 (1), p.34-41 |
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| source | MEDLINE; Free E-Journal (出版社公開部分のみ）; Oxford journals; Alma/SFX Local Collection |
| subjects | Bacteria Deoxyribonucleic acid DNA DNA, Bacterial - analysis Gene amplification Humans Infertility Medicine Methods Process controls RNA, Messenger - metabolism RNA, Ribosomal, 16S - metabolism Stem cells Stem Cells - metabolism |
| title | Sterility Testing of Stem Cell Products by Broad-Range Bacterial 16S Ribosomal DNA Polymerase Chain Reaction |
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