Evaluation of different real time PCRs for the detection of Pneumocystis jirovecii DNA in formalin-fixed paraffin-embedded bronchoalveolar lavage samples
The presence of Pneumocystis jirovecii in fresh clinical materials can be detected by PCR with high sensitivity and is thus preferred over microscopic methods. However, fresh materials are not always available, and on formalin-fixed paraffin-embedded materials, PCR may result in reduced detection ra...
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| Veröffentlicht in: | Experimental and molecular pathology 2015-06, Vol.98 (3), p.390-392 |
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| creator | de Leeuw, Bertie H.C.G.M. Voskuil, W. Sebastiaan Maraha, Boulos van der Zee, Anneke Westenend, Pieter J. Kusters, Johannes G. |
| description | The presence of Pneumocystis jirovecii in fresh clinical materials can be detected by PCR with high sensitivity and is thus preferred over microscopic methods. However, fresh materials are not always available, and on formalin-fixed paraffin-embedded materials, PCR may result in reduced detection rates. In this study the diagnostic sensitivity of P. jirovecii real time PCR on DNA isolated from fresh bronchoalveolar lavage (BAL) samples versus that from matched FFPE derived DNA is analyzed. Our results indicate that when targeting a small DNA fragment P. jirovecii PCR can be performed on FFPE BAL samples with acceptable sensitivity (up to 83.3%). This is considerably higher than the 33.3% positives observed by classical staining of these samples. |
| doi_str_mv | 10.1016/j.yexmp.2015.03.021 |
| format | Article |
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This is considerably higher than the 33.3% positives observed by classical staining of these samples.</description><identifier>ISSN: 0014-4800</identifier><identifier>EISSN: 1096-0945</identifier><identifier>DOI: 10.1016/j.yexmp.2015.03.021</identifier><identifier>PMID: 25779023</identifier><language>eng</language><publisher>Netherlands: Elsevier Inc</publisher><subject>Bronchoalveolar Lavage Fluid - microbiology ; DNA, Bacterial - genetics ; DNA, Bacterial - isolation & purification ; Formaldehyde - chemistry ; Formalin-fixed paraffin-embedded ; Humans ; Paraffin Embedding ; PCR ; Pneumocystis carinii - genetics ; Pneumocystis carinii - isolation & purification ; Pneumocystis jirovecii ; Real-Time Polymerase Chain Reaction - methods ; Sensitivity and Specificity ; Tissue Fixation</subject><ispartof>Experimental and molecular pathology, 2015-06, Vol.98 (3), p.390-392</ispartof><rights>2015 Elsevier Inc.</rights><rights>Copyright © 2015 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c359t-35686575e790b008109622ec2ece6b5420f030fe8c89cd937f5539b5d11b1ac23</citedby><cites>FETCH-LOGICAL-c359t-35686575e790b008109622ec2ece6b5420f030fe8c89cd937f5539b5d11b1ac23</cites><orcidid>0000-0001-7671-0736 ; 0000-0002-9967-6574</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.yexmp.2015.03.021$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25779023$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>de Leeuw, Bertie H.C.G.M.</creatorcontrib><creatorcontrib>Voskuil, W. Sebastiaan</creatorcontrib><creatorcontrib>Maraha, Boulos</creatorcontrib><creatorcontrib>van der Zee, Anneke</creatorcontrib><creatorcontrib>Westenend, Pieter J.</creatorcontrib><creatorcontrib>Kusters, Johannes G.</creatorcontrib><title>Evaluation of different real time PCRs for the detection of Pneumocystis jirovecii DNA in formalin-fixed paraffin-embedded bronchoalveolar lavage samples</title><title>Experimental and molecular pathology</title><addtitle>Exp Mol Pathol</addtitle><description>The presence of Pneumocystis jirovecii in fresh clinical materials can be detected by PCR with high sensitivity and is thus preferred over microscopic methods. However, fresh materials are not always available, and on formalin-fixed paraffin-embedded materials, PCR may result in reduced detection rates. In this study the diagnostic sensitivity of P. jirovecii real time PCR on DNA isolated from fresh bronchoalveolar lavage (BAL) samples versus that from matched FFPE derived DNA is analyzed. Our results indicate that when targeting a small DNA fragment P. jirovecii PCR can be performed on FFPE BAL samples with acceptable sensitivity (up to 83.3%). This is considerably higher than the 33.3% positives observed by classical staining of these samples.</description><subject>Bronchoalveolar Lavage Fluid - microbiology</subject><subject>DNA, Bacterial - genetics</subject><subject>DNA, Bacterial - isolation & purification</subject><subject>Formaldehyde - chemistry</subject><subject>Formalin-fixed paraffin-embedded</subject><subject>Humans</subject><subject>Paraffin Embedding</subject><subject>PCR</subject><subject>Pneumocystis carinii - genetics</subject><subject>Pneumocystis carinii - isolation & purification</subject><subject>Pneumocystis jirovecii</subject><subject>Real-Time Polymerase Chain Reaction - methods</subject><subject>Sensitivity and Specificity</subject><subject>Tissue Fixation</subject><issn>0014-4800</issn><issn>1096-0945</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kcFu1DAURS0EokPhC5CQl2wSnu04kyxYVNNCK1VQIVhbjv1MPXLiYCdR51P4WzJMyxLJkmXrXD9dH0LeMigZsPrDvjzgQz-WHJgsQZTA2TOyYdDWBbSVfE42AKwqqgbgjLzKeQ8ALTD-kpxxud22wMWG_L5adJj15ONAo6PWO4cJh4km1IFOvkd6t_uWqYuJTvdILU5onui7Aec-mkOefKZ7n-KCxnt6-eWC-uEY6XXwQ-H8A1o66qSdW4_Yd2jtetOlOJj7qMOCMehEg170T6RZ92PA_Jq8cDpkfPO4n5Mfn66-766L26-fb3YXt4URsp0KIeumlluJa6EOoDn25xzNurDuZMXBgQCHjWlaY1uxdVKKtpOWsY5pw8U5eX96d0zx14x5Ur3PBkPQA8Y5K1Y3jNesqusVFSfUpJhzQqfG5HudDoqBOjpRe_XXiTo6USDU6mRNvXscMHc92n-ZJwkr8PEE4Fpz8ZhUNh4Hg9an9bOVjf6_A_4ASzagww</recordid><startdate>201506</startdate><enddate>201506</enddate><creator>de Leeuw, Bertie H.C.G.M.</creator><creator>Voskuil, W. 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Sebastiaan ; Maraha, Boulos ; van der Zee, Anneke ; Westenend, Pieter J. ; Kusters, Johannes G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c359t-35686575e790b008109622ec2ece6b5420f030fe8c89cd937f5539b5d11b1ac23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Bronchoalveolar Lavage Fluid - microbiology</topic><topic>DNA, Bacterial - genetics</topic><topic>DNA, Bacterial - isolation & purification</topic><topic>Formaldehyde - chemistry</topic><topic>Formalin-fixed paraffin-embedded</topic><topic>Humans</topic><topic>Paraffin Embedding</topic><topic>PCR</topic><topic>Pneumocystis carinii - genetics</topic><topic>Pneumocystis carinii - isolation & purification</topic><topic>Pneumocystis jirovecii</topic><topic>Real-Time Polymerase Chain Reaction - methods</topic><topic>Sensitivity and Specificity</topic><topic>Tissue Fixation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>de Leeuw, Bertie H.C.G.M.</creatorcontrib><creatorcontrib>Voskuil, W. Sebastiaan</creatorcontrib><creatorcontrib>Maraha, Boulos</creatorcontrib><creatorcontrib>van der Zee, Anneke</creatorcontrib><creatorcontrib>Westenend, Pieter J.</creatorcontrib><creatorcontrib>Kusters, Johannes G.</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>Experimental and molecular pathology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>de Leeuw, Bertie H.C.G.M.</au><au>Voskuil, W. 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In this study the diagnostic sensitivity of P. jirovecii real time PCR on DNA isolated from fresh bronchoalveolar lavage (BAL) samples versus that from matched FFPE derived DNA is analyzed. Our results indicate that when targeting a small DNA fragment P. jirovecii PCR can be performed on FFPE BAL samples with acceptable sensitivity (up to 83.3%). This is considerably higher than the 33.3% positives observed by classical staining of these samples.</abstract><cop>Netherlands</cop><pub>Elsevier Inc</pub><pmid>25779023</pmid><doi>10.1016/j.yexmp.2015.03.021</doi><tpages>3</tpages><orcidid>https://orcid.org/0000-0001-7671-0736</orcidid><orcidid>https://orcid.org/0000-0002-9967-6574</orcidid></addata></record> |
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| ispartof | Experimental and molecular pathology, 2015-06, Vol.98 (3), p.390-392 |
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| language | eng |
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| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | Bronchoalveolar Lavage Fluid - microbiology DNA, Bacterial - genetics DNA, Bacterial - isolation & purification Formaldehyde - chemistry Formalin-fixed paraffin-embedded Humans Paraffin Embedding PCR Pneumocystis carinii - genetics Pneumocystis carinii - isolation & purification Pneumocystis jirovecii Real-Time Polymerase Chain Reaction - methods Sensitivity and Specificity Tissue Fixation |
| title | Evaluation of different real time PCRs for the detection of Pneumocystis jirovecii DNA in formalin-fixed paraffin-embedded bronchoalveolar lavage samples |
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