Detection and quantification of the iap gene of Listeria monocytogenes and Listeria innocua by a new real-time quantitative PCR assay
A real-time quantitative polymerase chain reaction (PCR) assay for direct detection and enumeration of Listeria monocytogenes and Listeria innocua was developed and applied to artificially contaminated milk samples. The iap gene present in both species was used as a target for amplification of a 175...
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| Veröffentlicht in: | Research in microbiology 2001-01, Vol.152 (1), p.37-46 |
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| creator | Hein, Ingeborg Klein, Dieter Lehner, Angelika Bubert, Andreas Brandl, Ernst Wagner, Martin |
| description | A real-time quantitative polymerase chain reaction (PCR) assay for direct detection and enumeration of
Listeria monocytogenes and
Listeria innocua was developed and applied to artificially contaminated milk samples. The
iap gene present in both species was used as a target for amplification of a 175-bp (
L. monocytogenes) and a 309-bp (
L. innocua) fragment. To ensure that
L. monocytogenes and
L. innocua are specifically detectable, tests were carried out using 42
L. monocytogenes strains and 33
L. innocua strains belonging to different serovars. Specificity was also confirmed using 22 bacterial strains not belonging to the genus
Listeria, including closely related bacteria. In addition to specificity, the reported assay is characterized by a wide dynamic range of quantification and a high sensitivity, as we could detect as few as six copies of the
iap gene per PCR using purified DNA as template. When applied to direct detection and quantification of
L. monocytogenes in milk, the more rapid real-time quantitative PCR assay was as sensitive as the traditional plate count method, but real-time quantitative PCR-derived
iap gene copy numbers were one to two logs higher than colony-forming units obtained by the plate count method. |
| doi_str_mv | 10.1016/S0923-2508(00)01166-9 |
| format | Article |
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Listeria monocytogenes and
Listeria innocua was developed and applied to artificially contaminated milk samples. The
iap gene present in both species was used as a target for amplification of a 175-bp (
L. monocytogenes) and a 309-bp (
L. innocua) fragment. To ensure that
L. monocytogenes and
L. innocua are specifically detectable, tests were carried out using 42
L. monocytogenes strains and 33
L. innocua strains belonging to different serovars. Specificity was also confirmed using 22 bacterial strains not belonging to the genus
Listeria, including closely related bacteria. In addition to specificity, the reported assay is characterized by a wide dynamic range of quantification and a high sensitivity, as we could detect as few as six copies of the
iap gene per PCR using purified DNA as template. When applied to direct detection and quantification of
L. monocytogenes in milk, the more rapid real-time quantitative PCR assay was as sensitive as the traditional plate count method, but real-time quantitative PCR-derived
iap gene copy numbers were one to two logs higher than colony-forming units obtained by the plate count method.</description><identifier>ISSN: 0923-2508</identifier><identifier>EISSN: 1769-7123</identifier><identifier>DOI: 10.1016/S0923-2508(00)01166-9</identifier><identifier>PMID: 11281324</identifier><language>eng</language><publisher>Paris: Elsevier SAS</publisher><subject>Aap gene ; Animals ; Bacterial Proteins - genetics ; Bacteriological methods and techniques used in bacteriology ; Bacteriology ; Biological and medical sciences ; Colony Count, Microbial ; Culture Media ; DNA Primers ; DNA, Bacterial - analysis ; DNA, Bacterial - isolation & purification ; Food industries ; Fundamental and applied biological sciences. Psychology ; Genes, Bacterial ; Listeria - genetics ; Listeria - isolation & purification ; Listeria innocua ; Listeria monocytogenes ; Listeria monocytogenes - genetics ; Listeria monocytogenes - growth & development ; Listeria monocytogenes - isolation & purification ; Microbiology ; milk ; Milk - microbiology ; Milk and cheese industries. Ice creams ; PCR ; Polymerase Chain Reaction - methods ; Sensitivity and Specificity</subject><ispartof>Research in microbiology, 2001-01, Vol.152 (1), p.37-46</ispartof><rights>2001 Éditions scientifiques et médicales Elsevier SAS</rights><rights>2001 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c533t-d3e5e903d8e146e2ce968d574014955c4a7458a15a8a448678b06f00090b76e03</citedby><cites>FETCH-LOGICAL-c533t-d3e5e903d8e146e2ce968d574014955c4a7458a15a8a448678b06f00090b76e03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0923250800011669$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27903,27904,65309</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=910776$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11281324$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hein, Ingeborg</creatorcontrib><creatorcontrib>Klein, Dieter</creatorcontrib><creatorcontrib>Lehner, Angelika</creatorcontrib><creatorcontrib>Bubert, Andreas</creatorcontrib><creatorcontrib>Brandl, Ernst</creatorcontrib><creatorcontrib>Wagner, Martin</creatorcontrib><title>Detection and quantification of the iap gene of Listeria monocytogenes and Listeria innocua by a new real-time quantitative PCR assay</title><title>Research in microbiology</title><addtitle>Res Microbiol</addtitle><description>A real-time quantitative polymerase chain reaction (PCR) assay for direct detection and enumeration of
Listeria monocytogenes and
Listeria innocua was developed and applied to artificially contaminated milk samples. The
iap gene present in both species was used as a target for amplification of a 175-bp (
L. monocytogenes) and a 309-bp (
L. innocua) fragment. To ensure that
L. monocytogenes and
L. innocua are specifically detectable, tests were carried out using 42
L. monocytogenes strains and 33
L. innocua strains belonging to different serovars. Specificity was also confirmed using 22 bacterial strains not belonging to the genus
Listeria, including closely related bacteria. In addition to specificity, the reported assay is characterized by a wide dynamic range of quantification and a high sensitivity, as we could detect as few as six copies of the
iap gene per PCR using purified DNA as template. When applied to direct detection and quantification of
L. monocytogenes in milk, the more rapid real-time quantitative PCR assay was as sensitive as the traditional plate count method, but real-time quantitative PCR-derived
iap gene copy numbers were one to two logs higher than colony-forming units obtained by the plate count method.</description><subject>Aap gene</subject><subject>Animals</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacteriological methods and techniques used in bacteriology</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Colony Count, Microbial</subject><subject>Culture Media</subject><subject>DNA Primers</subject><subject>DNA, Bacterial - analysis</subject><subject>DNA, Bacterial - isolation & purification</subject><subject>Food industries</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes, Bacterial</subject><subject>Listeria - genetics</subject><subject>Listeria - isolation & purification</subject><subject>Listeria innocua</subject><subject>Listeria monocytogenes</subject><subject>Listeria monocytogenes - genetics</subject><subject>Listeria monocytogenes - growth & development</subject><subject>Listeria monocytogenes - isolation & purification</subject><subject>Microbiology</subject><subject>milk</subject><subject>Milk - microbiology</subject><subject>Milk and cheese industries. Ice creams</subject><subject>PCR</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Sensitivity and Specificity</subject><issn>0923-2508</issn><issn>1769-7123</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkduKFDEQhoMo7uzqIygBQdaL1krn1LkSGY8woHi4Dpl0tUZ60rNJemUewPe2e6YZL_cqJP9XVaE-Qp4weMmAqVffwNS8qiU01wAvgDGlKnOPrJhWptKs5vfJ6oxckMucfwMwqbV4SC4YqxvGa7Eif99iQV_CEKmLLb0ZXSyhC94dn4aOll9Ig9vTnxhxvm9CLpiCo7shDv5QhjnIx-JzFOIUjY5uD9TRiH9oQtdXJexwGVCm9rdIv6y_UpezOzwiDzrXZ3y8nFfkx_t339cfq83nD5_WbzaVl5yXquUo0QBvG2RCYe3RqKaVWgATRkovnBaycUy6xgnRKN1sQXUAYGCrFQK_Is9PffdpuBkxF7sL2WPfu4jDmK3W04ok13eCTDdCcTOD8gT6NOScsLP7FHYuHSwDO4uyR1F2tmAB7FGUNVPd02XAuN1h-79qMTMBzxbAZe_6LrnoQz5zhoHWaqJenyictnYbMNnsA0aPbUiTVtsO4Y6P_AOXu683</recordid><startdate>20010101</startdate><enddate>20010101</enddate><creator>Hein, Ingeborg</creator><creator>Klein, Dieter</creator><creator>Lehner, Angelika</creator><creator>Bubert, Andreas</creator><creator>Brandl, Ernst</creator><creator>Wagner, Martin</creator><general>Elsevier SAS</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>7QL</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>20010101</creationdate><title>Detection and quantification of the iap gene of Listeria monocytogenes and Listeria innocua by a new real-time quantitative PCR assay</title><author>Hein, Ingeborg ; Klein, Dieter ; Lehner, Angelika ; Bubert, Andreas ; Brandl, Ernst ; Wagner, Martin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c533t-d3e5e903d8e146e2ce968d574014955c4a7458a15a8a448678b06f00090b76e03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Aap gene</topic><topic>Animals</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacteriological methods and techniques used in bacteriology</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Colony Count, Microbial</topic><topic>Culture Media</topic><topic>DNA Primers</topic><topic>DNA, Bacterial - analysis</topic><topic>DNA, Bacterial - isolation & purification</topic><topic>Food industries</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes, Bacterial</topic><topic>Listeria - genetics</topic><topic>Listeria - isolation & purification</topic><topic>Listeria innocua</topic><topic>Listeria monocytogenes</topic><topic>Listeria monocytogenes - genetics</topic><topic>Listeria monocytogenes - growth & development</topic><topic>Listeria monocytogenes - isolation & purification</topic><topic>Microbiology</topic><topic>milk</topic><topic>Milk - microbiology</topic><topic>Milk and cheese industries. Ice creams</topic><topic>PCR</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hein, Ingeborg</creatorcontrib><creatorcontrib>Klein, Dieter</creatorcontrib><creatorcontrib>Lehner, Angelika</creatorcontrib><creatorcontrib>Bubert, Andreas</creatorcontrib><creatorcontrib>Brandl, Ernst</creatorcontrib><creatorcontrib>Wagner, Martin</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>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Research in microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hein, Ingeborg</au><au>Klein, Dieter</au><au>Lehner, Angelika</au><au>Bubert, Andreas</au><au>Brandl, Ernst</au><au>Wagner, Martin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection and quantification of the iap gene of Listeria monocytogenes and Listeria innocua by a new real-time quantitative PCR assay</atitle><jtitle>Research in microbiology</jtitle><addtitle>Res Microbiol</addtitle><date>2001-01-01</date><risdate>2001</risdate><volume>152</volume><issue>1</issue><spage>37</spage><epage>46</epage><pages>37-46</pages><issn>0923-2508</issn><eissn>1769-7123</eissn><abstract>A real-time quantitative polymerase chain reaction (PCR) assay for direct detection and enumeration of
Listeria monocytogenes and
Listeria innocua was developed and applied to artificially contaminated milk samples. The
iap gene present in both species was used as a target for amplification of a 175-bp (
L. monocytogenes) and a 309-bp (
L. innocua) fragment. To ensure that
L. monocytogenes and
L. innocua are specifically detectable, tests were carried out using 42
L. monocytogenes strains and 33
L. innocua strains belonging to different serovars. Specificity was also confirmed using 22 bacterial strains not belonging to the genus
Listeria, including closely related bacteria. In addition to specificity, the reported assay is characterized by a wide dynamic range of quantification and a high sensitivity, as we could detect as few as six copies of the
iap gene per PCR using purified DNA as template. When applied to direct detection and quantification of
L. monocytogenes in milk, the more rapid real-time quantitative PCR assay was as sensitive as the traditional plate count method, but real-time quantitative PCR-derived
iap gene copy numbers were one to two logs higher than colony-forming units obtained by the plate count method.</abstract><cop>Paris</cop><pub>Elsevier SAS</pub><pmid>11281324</pmid><doi>10.1016/S0923-2508(00)01166-9</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| issn | 0923-2508 1769-7123 |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Aap gene Animals Bacterial Proteins - genetics Bacteriological methods and techniques used in bacteriology Bacteriology Biological and medical sciences Colony Count, Microbial Culture Media DNA Primers DNA, Bacterial - analysis DNA, Bacterial - isolation & purification Food industries Fundamental and applied biological sciences. Psychology Genes, Bacterial Listeria - genetics Listeria - isolation & purification Listeria innocua Listeria monocytogenes Listeria monocytogenes - genetics Listeria monocytogenes - growth & development Listeria monocytogenes - isolation & purification Microbiology milk Milk - microbiology Milk and cheese industries. Ice creams PCR Polymerase Chain Reaction - methods Sensitivity and Specificity |
| title | Detection and quantification of the iap gene of Listeria monocytogenes and Listeria innocua by a new real-time quantitative PCR assay |
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