Evaluation of a real-time PCR assay for detection of Bordetella pertussis and B. parapertussis in clinical samples
Institut für Hygiene und Laboratoriumsmedizin, Klinikum Krefeld, Lutherplatz 40, 47805 Krefeld, Germany Corresponding author: Professor C.H. Wirsing von König (e-mail: wvk_hyg{at}klinikum-krefeld.de ). Received 16 June 2000; revised version received 6 Oct. 2000; accepted 8 Nov. 2000. Abstract A real...
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| Veröffentlicht in: | Journal of medical microbiology 2001-05, Vol.50 (5), p.436-440 |
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| creator | KOSTERS, KATRIN RIFFELMANN, MARION VON KONIG, CARL HEINZ WIRSING |
| description | Institut für Hygiene und Laboratoriumsmedizin, Klinikum Krefeld, Lutherplatz 40, 47805 Krefeld, Germany
Corresponding author: Professor C.H. Wirsing von König (e-mail: wvk_hyg{at}klinikum-krefeld.de ).
Received 16 June 2000; revised version received 6 Oct. 2000; accepted 8 Nov. 2000.
Abstract
A real-time PCR assay based on the TaqMan ® technology was developed for the detection of Bordetella pertussis and B. parapertussis in clinical samples. The assay was evaluated with 182 specimens from 153 patients with and without symptoms of pertussis. The analytical sensitivity ranged from 0.1 to 10 cfu for B. pertussis and B. parapertussis , respectively, and diagnostic sensitivity was 94.1% when culture was used as a reference. No sample from a patient without symptoms of pertussis was positive in PCR. Twenty-four of 28 patients who were negative by culture and positive by PCR assay met the CDC clinical case definition for pertussis; the remaining four patients had paroxysms of shorter duration. Intra- and inter-assay variation were |
| doi_str_mv | 10.1099/0022-1317-50-5-436 |
| format | Article |
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Corresponding author: Professor C.H. Wirsing von König (e-mail: wvk_hyg{at}klinikum-krefeld.de ).
Received 16 June 2000; revised version received 6 Oct. 2000; accepted 8 Nov. 2000.
Abstract
A real-time PCR assay based on the TaqMan ® technology was developed for the detection of Bordetella pertussis and B. parapertussis in clinical samples. The assay was evaluated with 182 specimens from 153 patients with and without symptoms of pertussis. The analytical sensitivity ranged from 0.1 to 10 cfu for B. pertussis and B. parapertussis , respectively, and diagnostic sensitivity was 94.1% when culture was used as a reference. No sample from a patient without symptoms of pertussis was positive in PCR. Twenty-four of 28 patients who were negative by culture and positive by PCR assay met the CDC clinical case definition for pertussis; the remaining four patients had paroxysms of shorter duration. Intra- and inter-assay variation were <5% and results were available within 4 h.</description><identifier>ISSN: 0022-2615</identifier><identifier>EISSN: 1473-5644</identifier><identifier>DOI: 10.1099/0022-1317-50-5-436</identifier><identifier>PMID: 11339251</identifier><identifier>CODEN: JMMIAV</identifier><language>eng</language><publisher>Reading: Soc General Microbiol</publisher><subject>Adolescent ; Adult ; Aged ; Biological and medical sciences ; Bordetella - genetics ; Bordetella - isolation & purification ; Bordetella Infections - microbiology ; Bordetella Infections - pathology ; Bordetella pertussis - genetics ; Bordetella pertussis - isolation & purification ; Child ; Child, Preschool ; DNA, Bacterial - genetics ; Female ; Fundamental and applied biological sciences. Psychology ; Humans ; Infant ; Infant, Newborn ; Male ; Microbiology ; Middle Aged ; Polymerase Chain Reaction - methods ; Sensitivity and Specificity</subject><ispartof>Journal of medical microbiology, 2001-05, Vol.50 (5), p.436-440</ispartof><rights>2002 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c502t-b8f2fa1804116dab169a7b9f31564f29f7c1f78d8271e8e112de72ab897087283</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,3746,3747,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14162181$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11339251$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>KOSTERS, KATRIN</creatorcontrib><creatorcontrib>RIFFELMANN, MARION</creatorcontrib><creatorcontrib>VON KONIG, CARL HEINZ WIRSING</creatorcontrib><title>Evaluation of a real-time PCR assay for detection of Bordetella pertussis and B. parapertussis in clinical samples</title><title>Journal of medical microbiology</title><addtitle>J Med Microbiol</addtitle><description>Institut für Hygiene und Laboratoriumsmedizin, Klinikum Krefeld, Lutherplatz 40, 47805 Krefeld, Germany
Corresponding author: Professor C.H. Wirsing von König (e-mail: wvk_hyg{at}klinikum-krefeld.de ).
Received 16 June 2000; revised version received 6 Oct. 2000; accepted 8 Nov. 2000.
Abstract
A real-time PCR assay based on the TaqMan ® technology was developed for the detection of Bordetella pertussis and B. parapertussis in clinical samples. The assay was evaluated with 182 specimens from 153 patients with and without symptoms of pertussis. The analytical sensitivity ranged from 0.1 to 10 cfu for B. pertussis and B. parapertussis , respectively, and diagnostic sensitivity was 94.1% when culture was used as a reference. No sample from a patient without symptoms of pertussis was positive in PCR. Twenty-four of 28 patients who were negative by culture and positive by PCR assay met the CDC clinical case definition for pertussis; the remaining four patients had paroxysms of shorter duration. Intra- and inter-assay variation were <5% and results were available within 4 h.</description><subject>Adolescent</subject><subject>Adult</subject><subject>Aged</subject><subject>Biological and medical sciences</subject><subject>Bordetella - genetics</subject><subject>Bordetella - isolation & purification</subject><subject>Bordetella Infections - microbiology</subject><subject>Bordetella Infections - pathology</subject><subject>Bordetella pertussis - genetics</subject><subject>Bordetella pertussis - isolation & purification</subject><subject>Child</subject><subject>Child, Preschool</subject><subject>DNA, Bacterial - genetics</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Infant</subject><subject>Infant, Newborn</subject><subject>Male</subject><subject>Microbiology</subject><subject>Middle Aged</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Sensitivity and Specificity</subject><issn>0022-2615</issn><issn>1473-5644</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcluFDEQhi0EIkPgBTggX4CTg8u92D6SUVikSCAEZ6u6204c9Yarmyhvj5sZmCOnUpW-2v6fsZcgL0Ba-05KpQQUoEUlRSXKon7EdlDqQlR1WT5muz-AqqE6Y8-I7qQEXRT2KTsDyFFVsGPp6hf2Ky5xGvkUOPLksRdLHDz_uv_GkQgfeJgS7_zi27_Y5ZS2vO-Rzz4tK1EkjmPHLy_4jAlPxTjyto9jbLHnhMPce3rOngTsyb84xnP248PV9_0ncf3l4-f9-2vRVlItojFBBQQjS4C6wwZqi7qxoYD8XVA26BaCNp1RGrzxAKrzWmFjrJZGK1OcszeHuXOafq6eFjdEarejRz-t5DImrSrtf0Ew9aaXzqA6gG2aiJIPbk5xwPTgQLrNErcp7jZLXCVd5bIluenVcfraDL47tRw9yMDrI4CUZQoJxzbSiSuhVmA27u2Bu403t_cxeXfjxyHmW5o4ubth-LfyN_jtoS8</recordid><startdate>20010501</startdate><enddate>20010501</enddate><creator>KOSTERS, KATRIN</creator><creator>RIFFELMANN, MARION</creator><creator>VON KONIG, CARL HEINZ WIRSING</creator><general>Soc General Microbiol</general><general>Society for General Microbiology</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>7QL</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20010501</creationdate><title>Evaluation of a real-time PCR assay for detection of Bordetella pertussis and B. parapertussis in clinical samples</title><author>KOSTERS, KATRIN ; RIFFELMANN, MARION ; VON KONIG, CARL HEINZ WIRSING</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c502t-b8f2fa1804116dab169a7b9f31564f29f7c1f78d8271e8e112de72ab897087283</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Adolescent</topic><topic>Adult</topic><topic>Aged</topic><topic>Biological and medical sciences</topic><topic>Bordetella - genetics</topic><topic>Bordetella - isolation & purification</topic><topic>Bordetella Infections - microbiology</topic><topic>Bordetella Infections - pathology</topic><topic>Bordetella pertussis - genetics</topic><topic>Bordetella pertussis - isolation & purification</topic><topic>Child</topic><topic>Child, Preschool</topic><topic>DNA, Bacterial - genetics</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Infant</topic><topic>Infant, Newborn</topic><topic>Male</topic><topic>Microbiology</topic><topic>Middle Aged</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>KOSTERS, KATRIN</creatorcontrib><creatorcontrib>RIFFELMANN, MARION</creatorcontrib><creatorcontrib>VON KONIG, CARL HEINZ WIRSING</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of medical microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>KOSTERS, KATRIN</au><au>RIFFELMANN, MARION</au><au>VON KONIG, CARL HEINZ WIRSING</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Evaluation of a real-time PCR assay for detection of Bordetella pertussis and B. parapertussis in clinical samples</atitle><jtitle>Journal of medical microbiology</jtitle><addtitle>J Med Microbiol</addtitle><date>2001-05-01</date><risdate>2001</risdate><volume>50</volume><issue>5</issue><spage>436</spage><epage>440</epage><pages>436-440</pages><issn>0022-2615</issn><eissn>1473-5644</eissn><coden>JMMIAV</coden><abstract>Institut für Hygiene und Laboratoriumsmedizin, Klinikum Krefeld, Lutherplatz 40, 47805 Krefeld, Germany
Corresponding author: Professor C.H. Wirsing von König (e-mail: wvk_hyg{at}klinikum-krefeld.de ).
Received 16 June 2000; revised version received 6 Oct. 2000; accepted 8 Nov. 2000.
Abstract
A real-time PCR assay based on the TaqMan ® technology was developed for the detection of Bordetella pertussis and B. parapertussis in clinical samples. The assay was evaluated with 182 specimens from 153 patients with and without symptoms of pertussis. The analytical sensitivity ranged from 0.1 to 10 cfu for B. pertussis and B. parapertussis , respectively, and diagnostic sensitivity was 94.1% when culture was used as a reference. No sample from a patient without symptoms of pertussis was positive in PCR. Twenty-four of 28 patients who were negative by culture and positive by PCR assay met the CDC clinical case definition for pertussis; the remaining four patients had paroxysms of shorter duration. Intra- and inter-assay variation were <5% and results were available within 4 h.</abstract><cop>Reading</cop><pub>Soc General Microbiol</pub><pmid>11339251</pmid><doi>10.1099/0022-1317-50-5-436</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Adolescent Adult Aged Biological and medical sciences Bordetella - genetics Bordetella - isolation & purification Bordetella Infections - microbiology Bordetella Infections - pathology Bordetella pertussis - genetics Bordetella pertussis - isolation & purification Child Child, Preschool DNA, Bacterial - genetics Female Fundamental and applied biological sciences. Psychology Humans Infant Infant, Newborn Male Microbiology Middle Aged Polymerase Chain Reaction - methods Sensitivity and Specificity |
| title | Evaluation of a real-time PCR assay for detection of Bordetella pertussis and B. parapertussis in clinical samples |
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