An evaluation of PCR primer sets used for detection of Propionibacterium acnes in prostate tissue samples
BACKGROUND Multiple studies have now shown that Propionibacterium acnes can be cultured from post‐prostatectomy derived prostate tissue samples. In contrast, both universal eubacterial 16S rDNA PCR and P. acnes‐specific 16S rDNA PCR have failed to detect this organism at a frequency similar to that...
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| Veröffentlicht in: | The Prostate 2008-10, Vol.68 (14), p.1492-1495 |
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| creator | Sfanos, Karen S. Isaacs, William B. |
| description | BACKGROUND
Multiple studies have now shown that Propionibacterium acnes can be cultured from post‐prostatectomy derived prostate tissue samples. In contrast, both universal eubacterial 16S rDNA PCR and P. acnes‐specific 16S rDNA PCR have failed to detect this organism at a frequency similar to that of bacterial culture. A potential explanation for this discrepancy, proposed by Cohen et al., involves mismatches in 16S rDNA primer sets used for bacterial detection.
METHODS
The sensitivity of both a previously published P. acnes‐specific primer set containing a potential mismatch and a new primer set with no mismatches was determined. Both primer sets were used to interrogate two sets of DNA samples derived from post‐prostatectomy prostate tissues that differed in the level of sterile precautions maintained during tissue collection.
RESULTS
The number of P. acnes positive samples was associated with the sterility of the sample collection process. In all instances, positive samples were determined to reflect low cell numbers ( |
| doi_str_mv | 10.1002/pros.20820 |
| format | Article |
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Multiple studies have now shown that Propionibacterium acnes can be cultured from post‐prostatectomy derived prostate tissue samples. In contrast, both universal eubacterial 16S rDNA PCR and P. acnes‐specific 16S rDNA PCR have failed to detect this organism at a frequency similar to that of bacterial culture. A potential explanation for this discrepancy, proposed by Cohen et al., involves mismatches in 16S rDNA primer sets used for bacterial detection.
METHODS
The sensitivity of both a previously published P. acnes‐specific primer set containing a potential mismatch and a new primer set with no mismatches was determined. Both primer sets were used to interrogate two sets of DNA samples derived from post‐prostatectomy prostate tissues that differed in the level of sterile precautions maintained during tissue collection.
RESULTS
The number of P. acnes positive samples was associated with the sterility of the sample collection process. In all instances, positive samples were determined to reflect low cell numbers (<10 CFU).
CONCLUSIONS
Although the results of previous studies have shown that P. acnes is not the only organism potentially present in the prostates of prostate cancer patients, mismatches in PCR primer sets may have also influenced the sensitivity of P. acnes detection. When using PCR in determining the presence of P. acnes in the human prostate, care should be taken to establish the potential influence of exogenous contamination and, due to the sensitivity of the assay, samples exposed to the urethra during the collection process (prostatic secretions, TURP specimens) should not be used. Prostate 68: 1492–1495, 2008. © 2008 Wiley‐Liss, Inc.</description><identifier>ISSN: 0270-4137</identifier><identifier>EISSN: 1097-0045</identifier><identifier>DOI: 10.1002/pros.20820</identifier><identifier>PMID: 18651578</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>16S rDNA ; DNA Primers - chemistry ; DNA, Bacterial - chemistry ; DNA, Bacterial - genetics ; Humans ; Male ; Polymerase Chain Reaction - methods ; Propionibacterium acnes ; Propionibacterium acnes - genetics ; Propionibacterium acnes - isolation & purification ; Prostate - microbiology ; prostate tissue ; RNA, Ribosomal, 16S - chemistry ; RNA, Ribosomal, 16S - genetics ; Specimen Handling - methods</subject><ispartof>The Prostate, 2008-10, Vol.68 (14), p.1492-1495</ispartof><rights>Copyright © 2008 Wiley‐Liss, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4310-49f9e7276b5aebb63efb5bb589ab28cd97087a35f4af5cf4cec62961afbbbaba3</citedby><cites>FETCH-LOGICAL-c4310-49f9e7276b5aebb63efb5bb589ab28cd97087a35f4af5cf4cec62961afbbbaba3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fpros.20820$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fpros.20820$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27903,27904,45553,45554</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18651578$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sfanos, Karen S.</creatorcontrib><creatorcontrib>Isaacs, William B.</creatorcontrib><title>An evaluation of PCR primer sets used for detection of Propionibacterium acnes in prostate tissue samples</title><title>The Prostate</title><addtitle>Prostate</addtitle><description>BACKGROUND
Multiple studies have now shown that Propionibacterium acnes can be cultured from post‐prostatectomy derived prostate tissue samples. In contrast, both universal eubacterial 16S rDNA PCR and P. acnes‐specific 16S rDNA PCR have failed to detect this organism at a frequency similar to that of bacterial culture. A potential explanation for this discrepancy, proposed by Cohen et al., involves mismatches in 16S rDNA primer sets used for bacterial detection.
METHODS
The sensitivity of both a previously published P. acnes‐specific primer set containing a potential mismatch and a new primer set with no mismatches was determined. Both primer sets were used to interrogate two sets of DNA samples derived from post‐prostatectomy prostate tissues that differed in the level of sterile precautions maintained during tissue collection.
RESULTS
The number of P. acnes positive samples was associated with the sterility of the sample collection process. In all instances, positive samples were determined to reflect low cell numbers (<10 CFU).
CONCLUSIONS
Although the results of previous studies have shown that P. acnes is not the only organism potentially present in the prostates of prostate cancer patients, mismatches in PCR primer sets may have also influenced the sensitivity of P. acnes detection. When using PCR in determining the presence of P. acnes in the human prostate, care should be taken to establish the potential influence of exogenous contamination and, due to the sensitivity of the assay, samples exposed to the urethra during the collection process (prostatic secretions, TURP specimens) should not be used. Prostate 68: 1492–1495, 2008. © 2008 Wiley‐Liss, Inc.</description><subject>16S rDNA</subject><subject>DNA Primers - chemistry</subject><subject>DNA, Bacterial - chemistry</subject><subject>DNA, Bacterial - genetics</subject><subject>Humans</subject><subject>Male</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Propionibacterium acnes</subject><subject>Propionibacterium acnes - genetics</subject><subject>Propionibacterium acnes - isolation & purification</subject><subject>Prostate - microbiology</subject><subject>prostate tissue</subject><subject>RNA, Ribosomal, 16S - chemistry</subject><subject>RNA, Ribosomal, 16S - genetics</subject><subject>Specimen Handling - methods</subject><issn>0270-4137</issn><issn>1097-0045</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE9P3DAQxS1EBVvKpR-g8olDpdBxEsfJEUUtLUJloa3KzbK9Y8mQP4vHKfDtm-1u6a2nmcPvPb33GHsr4FQA5B_WcaTTHOoc9thCQKMygFLuswXkCrJSFOqQvSa6A5hxyA_YoagrKaSqFyycDRx_mW4yKYwDHz1ftjd8HUOPkRMm4hPhivsx8hUmdC9UHNfzG6xxCWOYem7cgMTDwDdxkknIUyCakJPp1x3SG_bKm47weHeP2I9PH7-3n7PLq_Mv7dll5spCzHEb36DKVWWlQWurAr2V1sq6MTav3apRUCtTSF8aL50vHboqbyphvLXWWFMcsZOt75zjYUJKug_ksOvMgONEumpK1dQAM_h-C7o5MEX0elPbxGctQG-G1Zsm-s-wM_xu5zrZHlf_0N2SMyC2wGPo8Pk_Vnp5c_Xtr2m21QRK-PSiMfFeV6pQUv_8eq7bC3XbXl9c62XxG1LMlaM</recordid><startdate>20081001</startdate><enddate>20081001</enddate><creator>Sfanos, Karen S.</creator><creator>Isaacs, William B.</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><scope>BSCLL</scope><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>20081001</creationdate><title>An evaluation of PCR primer sets used for detection of Propionibacterium acnes in prostate tissue samples</title><author>Sfanos, Karen S. ; Isaacs, William B.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4310-49f9e7276b5aebb63efb5bb589ab28cd97087a35f4af5cf4cec62961afbbbaba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>16S rDNA</topic><topic>DNA Primers - chemistry</topic><topic>DNA, Bacterial - chemistry</topic><topic>DNA, Bacterial - genetics</topic><topic>Humans</topic><topic>Male</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Propionibacterium acnes</topic><topic>Propionibacterium acnes - genetics</topic><topic>Propionibacterium acnes - isolation & purification</topic><topic>Prostate - microbiology</topic><topic>prostate tissue</topic><topic>RNA, Ribosomal, 16S - chemistry</topic><topic>RNA, Ribosomal, 16S - genetics</topic><topic>Specimen Handling - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sfanos, Karen S.</creatorcontrib><creatorcontrib>Isaacs, William B.</creatorcontrib><collection>Istex</collection><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>The Prostate</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sfanos, Karen S.</au><au>Isaacs, William B.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An evaluation of PCR primer sets used for detection of Propionibacterium acnes in prostate tissue samples</atitle><jtitle>The Prostate</jtitle><addtitle>Prostate</addtitle><date>2008-10-01</date><risdate>2008</risdate><volume>68</volume><issue>14</issue><spage>1492</spage><epage>1495</epage><pages>1492-1495</pages><issn>0270-4137</issn><eissn>1097-0045</eissn><abstract>BACKGROUND
Multiple studies have now shown that Propionibacterium acnes can be cultured from post‐prostatectomy derived prostate tissue samples. In contrast, both universal eubacterial 16S rDNA PCR and P. acnes‐specific 16S rDNA PCR have failed to detect this organism at a frequency similar to that of bacterial culture. A potential explanation for this discrepancy, proposed by Cohen et al., involves mismatches in 16S rDNA primer sets used for bacterial detection.
METHODS
The sensitivity of both a previously published P. acnes‐specific primer set containing a potential mismatch and a new primer set with no mismatches was determined. Both primer sets were used to interrogate two sets of DNA samples derived from post‐prostatectomy prostate tissues that differed in the level of sterile precautions maintained during tissue collection.
RESULTS
The number of P. acnes positive samples was associated with the sterility of the sample collection process. In all instances, positive samples were determined to reflect low cell numbers (<10 CFU).
CONCLUSIONS
Although the results of previous studies have shown that P. acnes is not the only organism potentially present in the prostates of prostate cancer patients, mismatches in PCR primer sets may have also influenced the sensitivity of P. acnes detection. When using PCR in determining the presence of P. acnes in the human prostate, care should be taken to establish the potential influence of exogenous contamination and, due to the sensitivity of the assay, samples exposed to the urethra during the collection process (prostatic secretions, TURP specimens) should not be used. Prostate 68: 1492–1495, 2008. © 2008 Wiley‐Liss, Inc.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>18651578</pmid><doi>10.1002/pros.20820</doi><tpages>4</tpages></addata></record> |
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| subjects | 16S rDNA DNA Primers - chemistry DNA, Bacterial - chemistry DNA, Bacterial - genetics Humans Male Polymerase Chain Reaction - methods Propionibacterium acnes Propionibacterium acnes - genetics Propionibacterium acnes - isolation & purification Prostate - microbiology prostate tissue RNA, Ribosomal, 16S - chemistry RNA, Ribosomal, 16S - genetics Specimen Handling - methods |
| title | An evaluation of PCR primer sets used for detection of Propionibacterium acnes in prostate tissue samples |
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