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An Improved Amplified Fragment Length Polymorphism (AFLP) Protocol for Discrimination of Listeria Isolates
Nine restriction enzyme combinations and 108 different primer combinations were initially tested for suitability for amplified fragment length polymorphism (AFLP) analysis of Listeria monocytogenes; the combination of Hin dIII and Hpy CH4IV showed consistently strong signals on gels, amplified an ad...
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| Published in: | Systematic and applied microbiology 2003-06, Vol.26 (2), p.236-244 |
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| creator | Keto-Timonen, Riikka O. Autio, Tiina J. Korkeala, Hannu J. |
| description | Nine restriction enzyme combinations and 108 different primer combinations were initially tested for suitability for amplified fragment length polymorphism (AFLP) analysis of
Listeria monocytogenes; the combination of
Hin dIII and
Hpy CH4IV showed consistently strong signals on gels, amplified an adequate number of DNA fragments and detected polymorphism among closely related strains based on
Asc I macrorestriction profiles. AFLP also distinguished between
L. monocytogenes, L. innocua, L. ivanovii,
L. seeligeri, L. welshimeri and
L. grayi species. All
Listeria species showed species-specific clusters, with less than 33% similarity between different species. A total of 34
L. monocytogenes strains were characterised by using both AFLP and pulsed-field gel electrophoresis (PFGE). The results of AFLP analysis of
L. monocytogenes strains were in concordance with those obtained by PFGE. Both methods identified 29 different genotypes of
L. monocytogenes and had a high discrimination index (>0.999). By combining the results of AFLP and PFGE, subtype discrimination was further improved. Numerical analysis of both AFLP and PFGE profiles yielded three genomic groups of
L. monocytogenes strains. AFLP was found to be faster and less labour-intensive than PFGE. We conclude that the AFLP protocol is a highly discriminatory, reproducible and valuable tool in characterisation of
Listeria strains and may also be suitable for
Listeria species identification. |
| doi_str_mv | 10.1078/072320203322346083 |
| format | article |
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Listeria monocytogenes; the combination of
Hin dIII and
Hpy CH4IV showed consistently strong signals on gels, amplified an adequate number of DNA fragments and detected polymorphism among closely related strains based on
Asc I macrorestriction profiles. AFLP also distinguished between
L. monocytogenes, L. innocua, L. ivanovii,
L. seeligeri, L. welshimeri and
L. grayi species. All
Listeria species showed species-specific clusters, with less than 33% similarity between different species. A total of 34
L. monocytogenes strains were characterised by using both AFLP and pulsed-field gel electrophoresis (PFGE). The results of AFLP analysis of
L. monocytogenes strains were in concordance with those obtained by PFGE. Both methods identified 29 different genotypes of
L. monocytogenes and had a high discrimination index (>0.999). By combining the results of AFLP and PFGE, subtype discrimination was further improved. Numerical analysis of both AFLP and PFGE profiles yielded three genomic groups of
L. monocytogenes strains. AFLP was found to be faster and less labour-intensive than PFGE. We conclude that the AFLP protocol is a highly discriminatory, reproducible and valuable tool in characterisation of
Listeria strains and may also be suitable for
Listeria species identification.</description><identifier>ISSN: 0723-2020</identifier><identifier>EISSN: 1618-0984</identifier><identifier>DOI: 10.1078/072320203322346083</identifier><identifier>PMID: 12866850</identifier><identifier>CODEN: SAMIDF</identifier><language>eng</language><publisher>Jena: Elsevier GmbH</publisher><subject>Amplified fragment length polymorphism (AFLP) ; Bacterial Typing Techniques - methods ; Bacteriology ; Biological and medical sciences ; DNA fingerprinting ; DNA Fingerprinting - methods ; DNA Restriction Enzymes ; DNA, Bacterial - genetics ; Electrophoresis, Gel, Pulsed-Field ; Fundamental and applied biological sciences. Psychology ; Genome, Bacterial ; Genotype ; Growth, nutrition, metabolism, transports, enzymes. Molecular biology ; Listeria - classification ; Listeria - genetics ; Listeria - isolation & purification ; Listeria monocytogenes ; Listeria monocytogenes - genetics ; Listeria monocytogenes - isolation & purification ; Listeria spp ; Microbiology ; Miscellaneous ; Mycology ; Nucleic Acid Amplification Techniques ; Polymorphism, Genetic ; pulsed-field gel electrophoresis (PFGE) ; Reproducibility of Results ; Species Specificity</subject><ispartof>Systematic and applied microbiology, 2003-06, Vol.26 (2), p.236-244</ispartof><rights>2003 Urban & Fischer Verlag</rights><rights>2004 INIST-CNRS</rights><rights>Copyright Urban & Fischer Verlag Jun 2003</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c440t-552c5d5e215b56b571ff4a297eea2b05e603ebc93e1e83e9e969ba1efa9d331c3</citedby><cites>FETCH-LOGICAL-c440t-552c5d5e215b56b571ff4a297eea2b05e603ebc93e1e83e9e969ba1efa9d331c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14888053$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12866850$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Keto-Timonen, Riikka O.</creatorcontrib><creatorcontrib>Autio, Tiina J.</creatorcontrib><creatorcontrib>Korkeala, Hannu J.</creatorcontrib><title>An Improved Amplified Fragment Length Polymorphism (AFLP) Protocol for Discrimination of Listeria Isolates</title><title>Systematic and applied microbiology</title><addtitle>Syst Appl Microbiol</addtitle><description>Nine restriction enzyme combinations and 108 different primer combinations were initially tested for suitability for amplified fragment length polymorphism (AFLP) analysis of
Listeria monocytogenes; the combination of
Hin dIII and
Hpy CH4IV showed consistently strong signals on gels, amplified an adequate number of DNA fragments and detected polymorphism among closely related strains based on
Asc I macrorestriction profiles. AFLP also distinguished between
L. monocytogenes, L. innocua, L. ivanovii,
L. seeligeri, L. welshimeri and
L. grayi species. All
Listeria species showed species-specific clusters, with less than 33% similarity between different species. A total of 34
L. monocytogenes strains were characterised by using both AFLP and pulsed-field gel electrophoresis (PFGE). The results of AFLP analysis of
L. monocytogenes strains were in concordance with those obtained by PFGE. Both methods identified 29 different genotypes of
L. monocytogenes and had a high discrimination index (>0.999). By combining the results of AFLP and PFGE, subtype discrimination was further improved. Numerical analysis of both AFLP and PFGE profiles yielded three genomic groups of
L. monocytogenes strains. AFLP was found to be faster and less labour-intensive than PFGE. We conclude that the AFLP protocol is a highly discriminatory, reproducible and valuable tool in characterisation of
Listeria strains and may also be suitable for
Listeria species identification.</description><subject>Amplified fragment length polymorphism (AFLP)</subject><subject>Bacterial Typing Techniques - methods</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>DNA fingerprinting</subject><subject>DNA Fingerprinting - methods</subject><subject>DNA Restriction Enzymes</subject><subject>DNA, Bacterial - genetics</subject><subject>Electrophoresis, Gel, Pulsed-Field</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genome, Bacterial</subject><subject>Genotype</subject><subject>Growth, nutrition, metabolism, transports, enzymes. 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Psychology</topic><topic>Genome, Bacterial</topic><topic>Genotype</topic><topic>Growth, nutrition, metabolism, transports, enzymes. Molecular biology</topic><topic>Listeria - classification</topic><topic>Listeria - genetics</topic><topic>Listeria - isolation & purification</topic><topic>Listeria monocytogenes</topic><topic>Listeria monocytogenes - genetics</topic><topic>Listeria monocytogenes - isolation & purification</topic><topic>Listeria spp</topic><topic>Microbiology</topic><topic>Miscellaneous</topic><topic>Mycology</topic><topic>Nucleic Acid Amplification Techniques</topic><topic>Polymorphism, Genetic</topic><topic>pulsed-field gel electrophoresis (PFGE)</topic><topic>Reproducibility of Results</topic><topic>Species Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Keto-Timonen, Riikka O.</creatorcontrib><creatorcontrib>Autio, Tiina J.</creatorcontrib><creatorcontrib>Korkeala, Hannu J.</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>ProQuest Central (Corporate)</collection><collection>Agricultural Science Collection</collection><collection>ProQuest Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>STEM Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>ProQuest Natural Science Collection</collection><collection>Earth, Atmospheric & Aquatic Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Agriculture Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>PML(ProQuest Medical Library)</collection><collection>Science Database (ProQuest)</collection><collection>Biological Science Database</collection><collection>Earth, Atmospheric & Aquatic Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>ProQuest Central Basic</collection><collection>SIRS Editorial</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Systematic and applied microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Keto-Timonen, Riikka O.</au><au>Autio, Tiina J.</au><au>Korkeala, Hannu J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An Improved Amplified Fragment Length Polymorphism (AFLP) Protocol for Discrimination of Listeria Isolates</atitle><jtitle>Systematic and applied microbiology</jtitle><addtitle>Syst Appl Microbiol</addtitle><date>2003-06-01</date><risdate>2003</risdate><volume>26</volume><issue>2</issue><spage>236</spage><epage>244</epage><pages>236-244</pages><issn>0723-2020</issn><eissn>1618-0984</eissn><coden>SAMIDF</coden><abstract>Nine restriction enzyme combinations and 108 different primer combinations were initially tested for suitability for amplified fragment length polymorphism (AFLP) analysis of
Listeria monocytogenes; the combination of
Hin dIII and
Hpy CH4IV showed consistently strong signals on gels, amplified an adequate number of DNA fragments and detected polymorphism among closely related strains based on
Asc I macrorestriction profiles. AFLP also distinguished between
L. monocytogenes, L. innocua, L. ivanovii,
L. seeligeri, L. welshimeri and
L. grayi species. All
Listeria species showed species-specific clusters, with less than 33% similarity between different species. A total of 34
L. monocytogenes strains were characterised by using both AFLP and pulsed-field gel electrophoresis (PFGE). The results of AFLP analysis of
L. monocytogenes strains were in concordance with those obtained by PFGE. Both methods identified 29 different genotypes of
L. monocytogenes and had a high discrimination index (>0.999). By combining the results of AFLP and PFGE, subtype discrimination was further improved. Numerical analysis of both AFLP and PFGE profiles yielded three genomic groups of
L. monocytogenes strains. AFLP was found to be faster and less labour-intensive than PFGE. We conclude that the AFLP protocol is a highly discriminatory, reproducible and valuable tool in characterisation of
Listeria strains and may also be suitable for
Listeria species identification.</abstract><cop>Jena</cop><pub>Elsevier GmbH</pub><pmid>12866850</pmid><doi>10.1078/072320203322346083</doi><tpages>9</tpages></addata></record> |
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| subjects | Amplified fragment length polymorphism (AFLP) Bacterial Typing Techniques - methods Bacteriology Biological and medical sciences DNA fingerprinting DNA Fingerprinting - methods DNA Restriction Enzymes DNA, Bacterial - genetics Electrophoresis, Gel, Pulsed-Field Fundamental and applied biological sciences. Psychology Genome, Bacterial Genotype Growth, nutrition, metabolism, transports, enzymes. Molecular biology Listeria - classification Listeria - genetics Listeria - isolation & purification Listeria monocytogenes Listeria monocytogenes - genetics Listeria monocytogenes - isolation & purification Listeria spp Microbiology Miscellaneous Mycology Nucleic Acid Amplification Techniques Polymorphism, Genetic pulsed-field gel electrophoresis (PFGE) Reproducibility of Results Species Specificity |
| title | An Improved Amplified Fragment Length Polymorphism (AFLP) Protocol for Discrimination of Listeria Isolates |
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