RNA-based Assay Demonstrated Enterococcus faecalis Metabolic Activity after Chemomechanical Procedures
Abstract Introduction Because ribosomal RNA (rRNA) indicates metabolic cell activity, this study aimed to evaluate the sensitivity of rRNA-based quantitative polymerase chain reaction (RT-qPCR) for the identification of active Enterococcus faecalis in root canals samples compared with a method based...
Gespeichert in:
| Veröffentlicht in: | Journal of endodontics 2015-09, Vol.41 (9), p.1441-1444 |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1444 |
|---|---|
| container_issue | 9 |
| container_start_page | 1441 |
| container_title | Journal of endodontics |
| container_volume | 41 |
| creator | Pinheiro, Ericka T., PhD Candeiro, George T., PhD Teixeira, Sílvia R., PhD Shin, Regina C., MSc Prado, Laís C., MSc Gavini, Giulio, PhD Mayer, Márcia P.A., PhD |
| description | Abstract Introduction Because ribosomal RNA (rRNA) indicates metabolic cell activity, this study aimed to evaluate the sensitivity of rRNA-based quantitative polymerase chain reaction (RT-qPCR) for the identification of active Enterococcus faecalis in root canals samples compared with a method based on ribosomal DNA (rDNA) (rRNA genes). Methods Samples were taken from 18 teeth with persistent/secondary intraradicular infection before (S1) and after (S2) chemomechanical preparation. RNA and DNA were extracted, and complementary DNA was synthesized from RNA using RT-PCR. Complementary DNA and genomic DNA were subjected to quantitative polymerase chain reaction with primers complementary for E. faecalis 16S rRNA sequence. Results E. faecalis was detected in 77.8% and 72.2% of S1 samples using rRNA- and rDNA-based assays, respectively. In contrast, E. faecalis was detected in only 33.3% of S2 samples using rDNA as the template compared with 61.1% using the rRNA-based method. The median concentration of rRNA copies of E. faecalis was significantly higher than rDNA copies, indicating a higher sensitivity for the method targeting rRNA in both S1 ( P |
| doi_str_mv | 10.1016/j.joen.2015.04.020 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1708896901</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>1_s2_0_S0099239915004070</els_id><sourcerecordid>1708896901</sourcerecordid><originalsourceid>FETCH-LOGICAL-c481t-ec4206bab3624415d04a6ffcade658af05054686634096b39eb6ec5a07cc2a8a3</originalsourceid><addsrcrecordid>eNp9kU2L1TAUhoMozp3RP-BCunTTevLZFkS4XMdRGD_wYx3S9JRJbZsxSQfuvzf1ji5cuAqcPO8L5zmEPKNQUaDq5ViNHpeKAZUViAoYPCA72tRNyaUUD8kOoG1Lxtv2jJzHOALQmvP6MTljCmqaP3dk-PJxX3YmYl_sYzTH4g3OfokpmJRHl0vC4K23do3FYNCaycXiAybT-cnZYm-Tu3PpWJghg8XhJodntDdmcRktPucs9mvA-IQ8GswU8en9e0G-v738dnhXXn-6en_YX5dWNDSVaAUD1ZmOKyYElT0Io4bBmh6VbMwAEqRQjVJcQKs63mKn0EoDtbXMNIZfkBen3tvgf64Yk55dtDhNZkG_Rk1raJpWtUAzyk6oDT7GgIO-DW424agp6M2vHvXmV29-NQid_ebQ8_v-tZux_xv5IzQDr04A5i3vHAYdrcMla3ABbdK9d__vf_1P3E7ut8wfeMQ4-jUs2Z-mOjIN-ut24e3AVAIIqIH_Ai_goeI</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1708896901</pqid></control><display><type>article</type><title>RNA-based Assay Demonstrated Enterococcus faecalis Metabolic Activity after Chemomechanical Procedures</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals</source><creator>Pinheiro, Ericka T., PhD ; Candeiro, George T., PhD ; Teixeira, Sílvia R., PhD ; Shin, Regina C., MSc ; Prado, Laís C., MSc ; Gavini, Giulio, PhD ; Mayer, Márcia P.A., PhD</creator><creatorcontrib>Pinheiro, Ericka T., PhD ; Candeiro, George T., PhD ; Teixeira, Sílvia R., PhD ; Shin, Regina C., MSc ; Prado, Laís C., MSc ; Gavini, Giulio, PhD ; Mayer, Márcia P.A., PhD</creatorcontrib><description>Abstract Introduction Because ribosomal RNA (rRNA) indicates metabolic cell activity, this study aimed to evaluate the sensitivity of rRNA-based quantitative polymerase chain reaction (RT-qPCR) for the identification of active Enterococcus faecalis in root canals samples compared with a method based on ribosomal DNA (rDNA) (rRNA genes). Methods Samples were taken from 18 teeth with persistent/secondary intraradicular infection before (S1) and after (S2) chemomechanical preparation. RNA and DNA were extracted, and complementary DNA was synthesized from RNA using RT-PCR. Complementary DNA and genomic DNA were subjected to quantitative polymerase chain reaction with primers complementary for E. faecalis 16S rRNA sequence. Results E. faecalis was detected in 77.8% and 72.2% of S1 samples using rRNA- and rDNA-based assays, respectively. In contrast, E. faecalis was detected in only 33.3% of S2 samples using rDNA as the template compared with 61.1% using the rRNA-based method. The median concentration of rRNA copies of E. faecalis was significantly higher than rDNA copies, indicating a higher sensitivity for the method targeting rRNA in both S1 ( P < .01) and S2 samples ( P < .05). After chemomechanical preparation, the number of rRNA and rDNA copies was significantly reduced ( P < .05). The high ratio of rRNA to rDNA copies in S2 samples suggested that active E. faecalis persisted in root canals after chemomechanical preparation. Conclusions The RT-qPCR assay provides a sensitive method for the identification of active E. faecalis from endodontic samples. Furthermore, the rRNA-based assay indicated that E. faecalis viable cells persisted in treated root canals, suggesting that it may be a useful tool for monitoring microbial load during endodontic treatment.</description><identifier>ISSN: 0099-2399</identifier><identifier>EISSN: 1878-3554</identifier><identifier>DOI: 10.1016/j.joen.2015.04.020</identifier><identifier>PMID: 26071099</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Dental Pulp Cavity - microbiology ; Dentistry ; DNA, Bacterial - analysis ; Endocrinology & Metabolism ; Endodontic ; Enterococcus faecalis ; Enterococcus faecalis - metabolism ; Humans ; quantitative polymerase chain reaction ; Real-Time Polymerase Chain Reaction ; ribosomal RNA–based polymerase chain reaction ; RNA, Ribosomal, 16S - analysis</subject><ispartof>Journal of endodontics, 2015-09, Vol.41 (9), p.1441-1444</ispartof><rights>American Association of Endodontists</rights><rights>2015 American Association of Endodontists</rights><rights>Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c481t-ec4206bab3624415d04a6ffcade658af05054686634096b39eb6ec5a07cc2a8a3</citedby><cites>FETCH-LOGICAL-c481t-ec4206bab3624415d04a6ffcade658af05054686634096b39eb6ec5a07cc2a8a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0099239915004070$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26071099$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pinheiro, Ericka T., PhD</creatorcontrib><creatorcontrib>Candeiro, George T., PhD</creatorcontrib><creatorcontrib>Teixeira, Sílvia R., PhD</creatorcontrib><creatorcontrib>Shin, Regina C., MSc</creatorcontrib><creatorcontrib>Prado, Laís C., MSc</creatorcontrib><creatorcontrib>Gavini, Giulio, PhD</creatorcontrib><creatorcontrib>Mayer, Márcia P.A., PhD</creatorcontrib><title>RNA-based Assay Demonstrated Enterococcus faecalis Metabolic Activity after Chemomechanical Procedures</title><title>Journal of endodontics</title><addtitle>J Endod</addtitle><description>Abstract Introduction Because ribosomal RNA (rRNA) indicates metabolic cell activity, this study aimed to evaluate the sensitivity of rRNA-based quantitative polymerase chain reaction (RT-qPCR) for the identification of active Enterococcus faecalis in root canals samples compared with a method based on ribosomal DNA (rDNA) (rRNA genes). Methods Samples were taken from 18 teeth with persistent/secondary intraradicular infection before (S1) and after (S2) chemomechanical preparation. RNA and DNA were extracted, and complementary DNA was synthesized from RNA using RT-PCR. Complementary DNA and genomic DNA were subjected to quantitative polymerase chain reaction with primers complementary for E. faecalis 16S rRNA sequence. Results E. faecalis was detected in 77.8% and 72.2% of S1 samples using rRNA- and rDNA-based assays, respectively. In contrast, E. faecalis was detected in only 33.3% of S2 samples using rDNA as the template compared with 61.1% using the rRNA-based method. The median concentration of rRNA copies of E. faecalis was significantly higher than rDNA copies, indicating a higher sensitivity for the method targeting rRNA in both S1 ( P < .01) and S2 samples ( P < .05). After chemomechanical preparation, the number of rRNA and rDNA copies was significantly reduced ( P < .05). The high ratio of rRNA to rDNA copies in S2 samples suggested that active E. faecalis persisted in root canals after chemomechanical preparation. Conclusions The RT-qPCR assay provides a sensitive method for the identification of active E. faecalis from endodontic samples. Furthermore, the rRNA-based assay indicated that E. faecalis viable cells persisted in treated root canals, suggesting that it may be a useful tool for monitoring microbial load during endodontic treatment.</description><subject>Dental Pulp Cavity - microbiology</subject><subject>Dentistry</subject><subject>DNA, Bacterial - analysis</subject><subject>Endocrinology & Metabolism</subject><subject>Endodontic</subject><subject>Enterococcus faecalis</subject><subject>Enterococcus faecalis - metabolism</subject><subject>Humans</subject><subject>quantitative polymerase chain reaction</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>ribosomal RNA–based polymerase chain reaction</subject><subject>RNA, Ribosomal, 16S - analysis</subject><issn>0099-2399</issn><issn>1878-3554</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU2L1TAUhoMozp3RP-BCunTTevLZFkS4XMdRGD_wYx3S9JRJbZsxSQfuvzf1ji5cuAqcPO8L5zmEPKNQUaDq5ViNHpeKAZUViAoYPCA72tRNyaUUD8kOoG1Lxtv2jJzHOALQmvP6MTljCmqaP3dk-PJxX3YmYl_sYzTH4g3OfokpmJRHl0vC4K23do3FYNCaycXiAybT-cnZYm-Tu3PpWJghg8XhJodntDdmcRktPucs9mvA-IQ8GswU8en9e0G-v738dnhXXn-6en_YX5dWNDSVaAUD1ZmOKyYElT0Io4bBmh6VbMwAEqRQjVJcQKs63mKn0EoDtbXMNIZfkBen3tvgf64Yk55dtDhNZkG_Rk1raJpWtUAzyk6oDT7GgIO-DW424agp6M2vHvXmV29-NQid_ebQ8_v-tZux_xv5IzQDr04A5i3vHAYdrcMla3ABbdK9d__vf_1P3E7ut8wfeMQ4-jUs2Z-mOjIN-ut24e3AVAIIqIH_Ai_goeI</recordid><startdate>20150901</startdate><enddate>20150901</enddate><creator>Pinheiro, Ericka T., PhD</creator><creator>Candeiro, George T., PhD</creator><creator>Teixeira, Sílvia R., PhD</creator><creator>Shin, Regina C., MSc</creator><creator>Prado, Laís C., MSc</creator><creator>Gavini, Giulio, PhD</creator><creator>Mayer, Márcia P.A., PhD</creator><general>Elsevier Inc</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>7X8</scope></search><sort><creationdate>20150901</creationdate><title>RNA-based Assay Demonstrated Enterococcus faecalis Metabolic Activity after Chemomechanical Procedures</title><author>Pinheiro, Ericka T., PhD ; Candeiro, George T., PhD ; Teixeira, Sílvia R., PhD ; Shin, Regina C., MSc ; Prado, Laís C., MSc ; Gavini, Giulio, PhD ; Mayer, Márcia P.A., PhD</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c481t-ec4206bab3624415d04a6ffcade658af05054686634096b39eb6ec5a07cc2a8a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Dental Pulp Cavity - microbiology</topic><topic>Dentistry</topic><topic>DNA, Bacterial - analysis</topic><topic>Endocrinology & Metabolism</topic><topic>Endodontic</topic><topic>Enterococcus faecalis</topic><topic>Enterococcus faecalis - metabolism</topic><topic>Humans</topic><topic>quantitative polymerase chain reaction</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>ribosomal RNA–based polymerase chain reaction</topic><topic>RNA, Ribosomal, 16S - analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pinheiro, Ericka T., PhD</creatorcontrib><creatorcontrib>Candeiro, George T., PhD</creatorcontrib><creatorcontrib>Teixeira, Sílvia R., PhD</creatorcontrib><creatorcontrib>Shin, Regina C., MSc</creatorcontrib><creatorcontrib>Prado, Laís C., MSc</creatorcontrib><creatorcontrib>Gavini, Giulio, PhD</creatorcontrib><creatorcontrib>Mayer, Márcia P.A., PhD</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of endodontics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pinheiro, Ericka T., PhD</au><au>Candeiro, George T., PhD</au><au>Teixeira, Sílvia R., PhD</au><au>Shin, Regina C., MSc</au><au>Prado, Laís C., MSc</au><au>Gavini, Giulio, PhD</au><au>Mayer, Márcia P.A., PhD</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>RNA-based Assay Demonstrated Enterococcus faecalis Metabolic Activity after Chemomechanical Procedures</atitle><jtitle>Journal of endodontics</jtitle><addtitle>J Endod</addtitle><date>2015-09-01</date><risdate>2015</risdate><volume>41</volume><issue>9</issue><spage>1441</spage><epage>1444</epage><pages>1441-1444</pages><issn>0099-2399</issn><eissn>1878-3554</eissn><abstract>Abstract Introduction Because ribosomal RNA (rRNA) indicates metabolic cell activity, this study aimed to evaluate the sensitivity of rRNA-based quantitative polymerase chain reaction (RT-qPCR) for the identification of active Enterococcus faecalis in root canals samples compared with a method based on ribosomal DNA (rDNA) (rRNA genes). Methods Samples were taken from 18 teeth with persistent/secondary intraradicular infection before (S1) and after (S2) chemomechanical preparation. RNA and DNA were extracted, and complementary DNA was synthesized from RNA using RT-PCR. Complementary DNA and genomic DNA were subjected to quantitative polymerase chain reaction with primers complementary for E. faecalis 16S rRNA sequence. Results E. faecalis was detected in 77.8% and 72.2% of S1 samples using rRNA- and rDNA-based assays, respectively. In contrast, E. faecalis was detected in only 33.3% of S2 samples using rDNA as the template compared with 61.1% using the rRNA-based method. The median concentration of rRNA copies of E. faecalis was significantly higher than rDNA copies, indicating a higher sensitivity for the method targeting rRNA in both S1 ( P < .01) and S2 samples ( P < .05). After chemomechanical preparation, the number of rRNA and rDNA copies was significantly reduced ( P < .05). The high ratio of rRNA to rDNA copies in S2 samples suggested that active E. faecalis persisted in root canals after chemomechanical preparation. Conclusions The RT-qPCR assay provides a sensitive method for the identification of active E. faecalis from endodontic samples. Furthermore, the rRNA-based assay indicated that E. faecalis viable cells persisted in treated root canals, suggesting that it may be a useful tool for monitoring microbial load during endodontic treatment.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>26071099</pmid><doi>10.1016/j.joen.2015.04.020</doi><tpages>4</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0099-2399 |
| ispartof | Journal of endodontics, 2015-09, Vol.41 (9), p.1441-1444 |
| issn | 0099-2399 1878-3554 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1708896901 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Dental Pulp Cavity - microbiology Dentistry DNA, Bacterial - analysis Endocrinology & Metabolism Endodontic Enterococcus faecalis Enterococcus faecalis - metabolism Humans quantitative polymerase chain reaction Real-Time Polymerase Chain Reaction ribosomal RNA–based polymerase chain reaction RNA, Ribosomal, 16S - analysis |
| title | RNA-based Assay Demonstrated Enterococcus faecalis Metabolic Activity after Chemomechanical Procedures |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-04T03%3A20%3A16IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=RNA-based%20Assay%20Demonstrated%20Enterococcus%20faecalis%20Metabolic%20Activity%20after%20Chemomechanical%20Procedures&rft.jtitle=Journal%20of%20endodontics&rft.au=Pinheiro,%20Ericka%20T.,%20PhD&rft.date=2015-09-01&rft.volume=41&rft.issue=9&rft.spage=1441&rft.epage=1444&rft.pages=1441-1444&rft.issn=0099-2399&rft.eissn=1878-3554&rft_id=info:doi/10.1016/j.joen.2015.04.020&rft_dat=%3Cproquest_cross%3E1708896901%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1708896901&rft_id=info:pmid/26071099&rft_els_id=1_s2_0_S0099239915004070&rfr_iscdi=true |