Accuracy of a routine real-time PCR assay for the diagnosis of Pneumocystis jirovecii pneumonia
Pneumocystis jirovecii is a common cause of life-threatening pneumonia among immunocompromised patients. Using 400 fresh bronchoalveolar lavage samples, we compared prospectively routine direct immunofluorescence assay (DFA) and a real-time PCR assay, performed on a LightCycler system, for the detec...
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| Veröffentlicht in: | Journal of microbiological methods 2008-10, Vol.75 (2), p.258-261 |
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| creator | Fillaux, Judith Malvy, Sonia Alvarez, Muriel Fabre, Richard Cassaing, Sophie Marchou, Bruno Linas, Marie-Denise Berry, Antoine |
| description | Pneumocystis jirovecii is a common cause of life-threatening pneumonia among immunocompromised patients. Using 400 fresh bronchoalveolar lavage samples, we compared prospectively routine direct immunofluorescence assay (DFA) and a real-time PCR assay, performed on a LightCycler system, for the detection of
P. jirovecii. Among the 66 PCR positive samples, 31 were positive by DFA. No patient was found as having the pattern “PCR−−ve/DFA+ve”. The semi-quantification of the
P. jirovecii DNA was represented by the cycle threshold (Ct). Using DFA as the gold standard, the sensitivity of the PCR was 100% for Ct
≥
28 and the specificity was 100% for Ct
<
22. Between these two points, the results could be discrepant. The patients of the “22
≤
Ct
<
28” group presented more frequently with a radiological interstitial syndrome than the “Ct
≥
28” group, and presented less frequently with HIV-infection and elevated lactate dehydrogenase (LDH) assay than in the “Ct
<
22” group. A negative PCR allowed us to exclude the
P. jirovecii pneumonia. The real-time PCR assay seems to be an accurate diagnosis method and could replace the DFA. The semi-quantitative results should be helpful to distinguish colonized, subclinically infected and
P. jirovecii pneumonia patients. |
| doi_str_mv | 10.1016/j.mimet.2008.06.009 |
| format | Article |
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P. jirovecii. Among the 66 PCR positive samples, 31 were positive by DFA. No patient was found as having the pattern “PCR−−ve/DFA+ve”. The semi-quantification of the
P. jirovecii DNA was represented by the cycle threshold (Ct). Using DFA as the gold standard, the sensitivity of the PCR was 100% for Ct
≥
28 and the specificity was 100% for Ct
<
22. Between these two points, the results could be discrepant. The patients of the “22
≤
Ct
<
28” group presented more frequently with a radiological interstitial syndrome than the “Ct
≥
28” group, and presented less frequently with HIV-infection and elevated lactate dehydrogenase (LDH) assay than in the “Ct
<
22” group. A negative PCR allowed us to exclude the
P. jirovecii pneumonia. The real-time PCR assay seems to be an accurate diagnosis method and could replace the DFA. The semi-quantitative results should be helpful to distinguish colonized, subclinically infected and
P. jirovecii pneumonia patients.</description><identifier>ISSN: 0167-7012</identifier><identifier>EISSN: 1872-8359</identifier><identifier>DOI: 10.1016/j.mimet.2008.06.009</identifier><identifier>PMID: 18606198</identifier><identifier>CODEN: JMIMDQ</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Aged ; Biological and medical sciences ; Bronchoalveolar Lavage Fluid - microbiology ; Diagnosis ; DNA Primers ; DNA, Bacterial - analysis ; Female ; Fluorescent Antibody Technique, Direct ; Fundamental and applied biological sciences. Psychology ; Human bronchoalveolar lavage ; Human immunodeficiency virus ; Humans ; Male ; Microbiology ; Middle Aged ; Mycological methods and techniques used in mycology ; Mycology ; Pathogenicity, host-agent relations, miscellaneous strains, epidemiology ; Pneumocystis ; Pneumocystis carinii - genetics ; Pneumocystis carinii - isolation & purification ; Pneumocystis jirovecii ; Pneumonia, Pneumocystis - diagnosis ; Pneumonia, Pneumocystis - microbiology ; Polymerase Chain Reaction - methods ; Real-time PCR ; Sensitivity and Specificity</subject><ispartof>Journal of microbiological methods, 2008-10, Vol.75 (2), p.258-261</ispartof><rights>2008 Elsevier B.V.</rights><rights>2009 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c418t-6b224413082f6ee96508c039ba3bf96d97e7c548409f2d47c5255fc936abd0513</citedby><cites>FETCH-LOGICAL-c418t-6b224413082f6ee96508c039ba3bf96d97e7c548409f2d47c5255fc936abd0513</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.mimet.2008.06.009$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>315,781,785,3551,27929,27930,46000</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=20708001$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18606198$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fillaux, Judith</creatorcontrib><creatorcontrib>Malvy, Sonia</creatorcontrib><creatorcontrib>Alvarez, Muriel</creatorcontrib><creatorcontrib>Fabre, Richard</creatorcontrib><creatorcontrib>Cassaing, Sophie</creatorcontrib><creatorcontrib>Marchou, Bruno</creatorcontrib><creatorcontrib>Linas, Marie-Denise</creatorcontrib><creatorcontrib>Berry, Antoine</creatorcontrib><title>Accuracy of a routine real-time PCR assay for the diagnosis of Pneumocystis jirovecii pneumonia</title><title>Journal of microbiological methods</title><addtitle>J Microbiol Methods</addtitle><description>Pneumocystis jirovecii is a common cause of life-threatening pneumonia among immunocompromised patients. Using 400 fresh bronchoalveolar lavage samples, we compared prospectively routine direct immunofluorescence assay (DFA) and a real-time PCR assay, performed on a LightCycler system, for the detection of
P. jirovecii. Among the 66 PCR positive samples, 31 were positive by DFA. No patient was found as having the pattern “PCR−−ve/DFA+ve”. The semi-quantification of the
P. jirovecii DNA was represented by the cycle threshold (Ct). Using DFA as the gold standard, the sensitivity of the PCR was 100% for Ct
≥
28 and the specificity was 100% for Ct
<
22. Between these two points, the results could be discrepant. The patients of the “22
≤
Ct
<
28” group presented more frequently with a radiological interstitial syndrome than the “Ct
≥
28” group, and presented less frequently with HIV-infection and elevated lactate dehydrogenase (LDH) assay than in the “Ct
<
22” group. A negative PCR allowed us to exclude the
P. jirovecii pneumonia. The real-time PCR assay seems to be an accurate diagnosis method and could replace the DFA. The semi-quantitative results should be helpful to distinguish colonized, subclinically infected and
P. jirovecii pneumonia patients.</description><subject>Aged</subject><subject>Biological and medical sciences</subject><subject>Bronchoalveolar Lavage Fluid - microbiology</subject><subject>Diagnosis</subject><subject>DNA Primers</subject><subject>DNA, Bacterial - analysis</subject><subject>Female</subject><subject>Fluorescent Antibody Technique, Direct</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Human bronchoalveolar lavage</subject><subject>Human immunodeficiency virus</subject><subject>Humans</subject><subject>Male</subject><subject>Microbiology</subject><subject>Middle Aged</subject><subject>Mycological methods and techniques used in mycology</subject><subject>Mycology</subject><subject>Pathogenicity, host-agent relations, miscellaneous strains, epidemiology</subject><subject>Pneumocystis</subject><subject>Pneumocystis carinii - genetics</subject><subject>Pneumocystis carinii - isolation & purification</subject><subject>Pneumocystis jirovecii</subject><subject>Pneumonia, Pneumocystis - diagnosis</subject><subject>Pneumonia, Pneumocystis - microbiology</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Real-time PCR</subject><subject>Sensitivity and Specificity</subject><issn>0167-7012</issn><issn>1872-8359</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1u1DAUhS0EokPhCZCQN7BLuLZjx16wqEb8SZVaoXZtOc4NeJTEg51UmrfH0xnBjq78o-8cXd2PkLcMagZMfdzVU5hwqTmArkHVAOYZ2TDd8koLaZ6TTaHaqgXGL8irnHcATIpGvyQXTCtQzOgNsVfer8n5A40DdTTFdQkz0oRurJZST2-3P6jL2R3oEBNdfiHtg_s5xxzyMXI74zpFf8hLee9Cig_oQ6D7x-85uNfkxeDGjG_O5yW5__L5bvutur75-n17dV35humlUh3nTcMEaD4oRKMkaA_CdE50g1G9abH1stENmIH3TblzKQdvhHJdD5KJS_Lh1LtP8feKebFTyB7H0c0Y12yVkZxpEE-CzIgySSsLKE6gTzHnhIPdpzC5dLAM7FGA3dlHAfYowIKyRUBJvTvXr92E_b_MeeMFeH8GXPZuHJKbfch_OQ4t6CKqcJ9OHJatPQRMNvuAs8c-JPSL7WP47yB_AP0RpGs</recordid><startdate>20081001</startdate><enddate>20081001</enddate><creator>Fillaux, Judith</creator><creator>Malvy, Sonia</creator><creator>Alvarez, Muriel</creator><creator>Fabre, Richard</creator><creator>Cassaing, Sophie</creator><creator>Marchou, Bruno</creator><creator>Linas, Marie-Denise</creator><creator>Berry, Antoine</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20081001</creationdate><title>Accuracy of a routine real-time PCR assay for the diagnosis of Pneumocystis jirovecii pneumonia</title><author>Fillaux, Judith ; Malvy, Sonia ; Alvarez, Muriel ; Fabre, Richard ; Cassaing, Sophie ; Marchou, Bruno ; Linas, Marie-Denise ; Berry, Antoine</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c418t-6b224413082f6ee96508c039ba3bf96d97e7c548409f2d47c5255fc936abd0513</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Aged</topic><topic>Biological and medical sciences</topic><topic>Bronchoalveolar Lavage Fluid - microbiology</topic><topic>Diagnosis</topic><topic>DNA Primers</topic><topic>DNA, Bacterial - analysis</topic><topic>Female</topic><topic>Fluorescent Antibody Technique, Direct</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Human bronchoalveolar lavage</topic><topic>Human immunodeficiency virus</topic><topic>Humans</topic><topic>Male</topic><topic>Microbiology</topic><topic>Middle Aged</topic><topic>Mycological methods and techniques used in mycology</topic><topic>Mycology</topic><topic>Pathogenicity, host-agent relations, miscellaneous strains, epidemiology</topic><topic>Pneumocystis</topic><topic>Pneumocystis carinii - genetics</topic><topic>Pneumocystis carinii - isolation & purification</topic><topic>Pneumocystis jirovecii</topic><topic>Pneumonia, Pneumocystis - diagnosis</topic><topic>Pneumonia, Pneumocystis - microbiology</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Real-time PCR</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Fillaux, Judith</creatorcontrib><creatorcontrib>Malvy, Sonia</creatorcontrib><creatorcontrib>Alvarez, Muriel</creatorcontrib><creatorcontrib>Fabre, Richard</creatorcontrib><creatorcontrib>Cassaing, Sophie</creatorcontrib><creatorcontrib>Marchou, Bruno</creatorcontrib><creatorcontrib>Linas, Marie-Denise</creatorcontrib><creatorcontrib>Berry, Antoine</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of microbiological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Fillaux, Judith</au><au>Malvy, Sonia</au><au>Alvarez, Muriel</au><au>Fabre, Richard</au><au>Cassaing, Sophie</au><au>Marchou, Bruno</au><au>Linas, Marie-Denise</au><au>Berry, Antoine</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Accuracy of a routine real-time PCR assay for the diagnosis of Pneumocystis jirovecii pneumonia</atitle><jtitle>Journal of microbiological methods</jtitle><addtitle>J Microbiol Methods</addtitle><date>2008-10-01</date><risdate>2008</risdate><volume>75</volume><issue>2</issue><spage>258</spage><epage>261</epage><pages>258-261</pages><issn>0167-7012</issn><eissn>1872-8359</eissn><coden>JMIMDQ</coden><abstract>Pneumocystis jirovecii is a common cause of life-threatening pneumonia among immunocompromised patients. Using 400 fresh bronchoalveolar lavage samples, we compared prospectively routine direct immunofluorescence assay (DFA) and a real-time PCR assay, performed on a LightCycler system, for the detection of
P. jirovecii. Among the 66 PCR positive samples, 31 were positive by DFA. No patient was found as having the pattern “PCR−−ve/DFA+ve”. The semi-quantification of the
P. jirovecii DNA was represented by the cycle threshold (Ct). Using DFA as the gold standard, the sensitivity of the PCR was 100% for Ct
≥
28 and the specificity was 100% for Ct
<
22. Between these two points, the results could be discrepant. The patients of the “22
≤
Ct
<
28” group presented more frequently with a radiological interstitial syndrome than the “Ct
≥
28” group, and presented less frequently with HIV-infection and elevated lactate dehydrogenase (LDH) assay than in the “Ct
<
22” group. A negative PCR allowed us to exclude the
P. jirovecii pneumonia. The real-time PCR assay seems to be an accurate diagnosis method and could replace the DFA. The semi-quantitative results should be helpful to distinguish colonized, subclinically infected and
P. jirovecii pneumonia patients.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>18606198</pmid><doi>10.1016/j.mimet.2008.06.009</doi><tpages>4</tpages></addata></record> |
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| subjects | Aged Biological and medical sciences Bronchoalveolar Lavage Fluid - microbiology Diagnosis DNA Primers DNA, Bacterial - analysis Female Fluorescent Antibody Technique, Direct Fundamental and applied biological sciences. Psychology Human bronchoalveolar lavage Human immunodeficiency virus Humans Male Microbiology Middle Aged Mycological methods and techniques used in mycology Mycology Pathogenicity, host-agent relations, miscellaneous strains, epidemiology Pneumocystis Pneumocystis carinii - genetics Pneumocystis carinii - isolation & purification Pneumocystis jirovecii Pneumonia, Pneumocystis - diagnosis Pneumonia, Pneumocystis - microbiology Polymerase Chain Reaction - methods Real-time PCR Sensitivity and Specificity |
| title | Accuracy of a routine real-time PCR assay for the diagnosis of Pneumocystis jirovecii pneumonia |
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