Specific detection and differentiation of two subspecies of Fusobacterium necrophorum by PCR
The phylogenic relationships of two subspecies of Fusobacterium necrophorum were investigated by randomly amplified polymorphism DNA–polymerase chain reaction (RAPD–PCR). With each of the 12 random primers, the DNA fingerprints generated were subjected to cluster analysis for dendrograms. The analys...
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| Veröffentlicht in: | Veterinary microbiology 2003-02, Vol.91 (2), p.183-195 |
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| creator | Narongwanichgarn, Wonganun Misawa, Naoaki Jin, Jing Hua Amoako, Kingsley Kwaku Kawaguchi, Emi Shinjo, Toshiharu Haga, Takeshi Goto, Yoshitaka |
| description | The phylogenic relationships of two subspecies of
Fusobacterium necrophorum were investigated by randomly amplified polymorphism DNA–polymerase chain reaction (RAPD–PCR). With each of the 12 random primers, the DNA fingerprints generated were subjected to cluster analysis for dendrograms. The analysis indicated that twelve strains were organized into two major clusters, and that all strains of each subspecies were confined to one cluster. Furthermore, two of the random primers examined each generated a unique band in
F. n. necrophorum strains. We cloned these specific bands and determined the nucleotide sequences. A search for amino acid sequence homologies revealed that the two specific fragments had significant homology to the
rpoB gene of
Lactococcus lactis subsp.
lactis and the hemagglutinin-related protein gene of
Ralstonia solanacearum, respectively. New specific primers designed for the
rpoB gene were able to amplify 900
bp fragments from both subspecies. However, the specific primers designed for the hemagglutinin-related protein gene amplified only a 250
bp fragment of the genome of the
F. n. necrophorum strains, suggesting that this gene is unique to
F. n. necrophorum. These results were further confirmed by dot blot hybridization. Finally, a one-step duplex PCR technique in a single tube for the rapid detection and differentiation of the
F. necrophorum subspecies was developed. |
| doi_str_mv | 10.1016/S0378-1135(02)00295-X |
| format | Article |
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Fusobacterium necrophorum were investigated by randomly amplified polymorphism DNA–polymerase chain reaction (RAPD–PCR). With each of the 12 random primers, the DNA fingerprints generated were subjected to cluster analysis for dendrograms. The analysis indicated that twelve strains were organized into two major clusters, and that all strains of each subspecies were confined to one cluster. Furthermore, two of the random primers examined each generated a unique band in
F. n. necrophorum strains. We cloned these specific bands and determined the nucleotide sequences. A search for amino acid sequence homologies revealed that the two specific fragments had significant homology to the
rpoB gene of
Lactococcus lactis subsp.
lactis and the hemagglutinin-related protein gene of
Ralstonia solanacearum, respectively. New specific primers designed for the
rpoB gene were able to amplify 900
bp fragments from both subspecies. However, the specific primers designed for the hemagglutinin-related protein gene amplified only a 250
bp fragment of the genome of the
F. n. necrophorum strains, suggesting that this gene is unique to
F. n. necrophorum. These results were further confirmed by dot blot hybridization. Finally, a one-step duplex PCR technique in a single tube for the rapid detection and differentiation of the
F. necrophorum subspecies was developed.</description><identifier>ISSN: 0378-1135</identifier><identifier>EISSN: 1873-2542</identifier><identifier>DOI: 10.1016/S0378-1135(02)00295-X</identifier><identifier>PMID: 12458167</identifier><identifier>CODEN: VMICDQ</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Bacteriological methods and techniques used in bacteriology ; Bacteriology ; Base Sequence ; Biological and medical sciences ; Cloning, Molecular ; Cluster Analysis ; DNA Primers - chemistry ; DNA Primers - genetics ; DNA, Bacterial - chemistry ; DNA, Bacterial - genetics ; Fundamental and applied biological sciences. Psychology ; Fusobacterium necrophorum ; Fusobacterium necrophorum - chemistry ; Fusobacterium necrophorum - classification ; Fusobacterium necrophorum - genetics ; Microbiology ; Molecular Sequence Data ; Nucleic Acid Hybridization ; PCR ; Phylogeny ; Polymerase Chain Reaction - veterinary ; Random Amplified Polymorphic DNA Technique - veterinary ; Sequence Analysis, DNA ; Specific detection and differentiation</subject><ispartof>Veterinary microbiology, 2003-02, Vol.91 (2), p.183-195</ispartof><rights>2002 Elsevier Science B.V.</rights><rights>2003 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c488t-1472cf1bc58bf2a1923e20b0fd78264e99f2eb0c0a4d751783e597f9a50c62f73</citedby><cites>FETCH-LOGICAL-c488t-1472cf1bc58bf2a1923e20b0fd78264e99f2eb0c0a4d751783e597f9a50c62f73</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S037811350200295X$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14408726$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12458167$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Narongwanichgarn, Wonganun</creatorcontrib><creatorcontrib>Misawa, Naoaki</creatorcontrib><creatorcontrib>Jin, Jing Hua</creatorcontrib><creatorcontrib>Amoako, Kingsley Kwaku</creatorcontrib><creatorcontrib>Kawaguchi, Emi</creatorcontrib><creatorcontrib>Shinjo, Toshiharu</creatorcontrib><creatorcontrib>Haga, Takeshi</creatorcontrib><creatorcontrib>Goto, Yoshitaka</creatorcontrib><title>Specific detection and differentiation of two subspecies of Fusobacterium necrophorum by PCR</title><title>Veterinary microbiology</title><addtitle>Vet Microbiol</addtitle><description>The phylogenic relationships of two subspecies of
Fusobacterium necrophorum were investigated by randomly amplified polymorphism DNA–polymerase chain reaction (RAPD–PCR). With each of the 12 random primers, the DNA fingerprints generated were subjected to cluster analysis for dendrograms. The analysis indicated that twelve strains were organized into two major clusters, and that all strains of each subspecies were confined to one cluster. Furthermore, two of the random primers examined each generated a unique band in
F. n. necrophorum strains. We cloned these specific bands and determined the nucleotide sequences. A search for amino acid sequence homologies revealed that the two specific fragments had significant homology to the
rpoB gene of
Lactococcus lactis subsp.
lactis and the hemagglutinin-related protein gene of
Ralstonia solanacearum, respectively. New specific primers designed for the
rpoB gene were able to amplify 900
bp fragments from both subspecies. However, the specific primers designed for the hemagglutinin-related protein gene amplified only a 250
bp fragment of the genome of the
F. n. necrophorum strains, suggesting that this gene is unique to
F. n. necrophorum. These results were further confirmed by dot blot hybridization. Finally, a one-step duplex PCR technique in a single tube for the rapid detection and differentiation of the
F. necrophorum subspecies was developed.</description><subject>Bacteriological methods and techniques used in bacteriology</subject><subject>Bacteriology</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cloning, Molecular</subject><subject>Cluster Analysis</subject><subject>DNA Primers - chemistry</subject><subject>DNA Primers - genetics</subject><subject>DNA, Bacterial - chemistry</subject><subject>DNA, Bacterial - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fusobacterium necrophorum</subject><subject>Fusobacterium necrophorum - chemistry</subject><subject>Fusobacterium necrophorum - classification</subject><subject>Fusobacterium necrophorum - genetics</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Nucleic Acid Hybridization</subject><subject>PCR</subject><subject>Phylogeny</subject><subject>Polymerase Chain Reaction - veterinary</subject><subject>Random Amplified Polymorphic DNA Technique - veterinary</subject><subject>Sequence Analysis, DNA</subject><subject>Specific detection and differentiation</subject><issn>0378-1135</issn><issn>1873-2542</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkV1rFTEQhkNR7LH6E5S9UfRidTKbbLJXIgerQsHSD-hFIWSzE5pyzuaY7Cr99-Z8YC97lWF43snkCWNvOHziwNvPl9AoXXPeyA-AHwGwk_XNEVtwrZoapcBnbPEfOWYvc74HANG18IIdcxRS81Yt2O3lhlzwwVUDTeSmEMfKjkM1BO8p0TgFu-tFX01_Y5XnPm8DlLed0znH3rqJUpjX1Uguxc1dTKXuH6rz5cUr9tzbVabXh_OEXZ9-u1r-qM9-ff-5_HpWO6H1VHOh0HneO6l7j5Z32BBCD35QGltBXeeRenBgxaAkV7oh2SnfWQmuRa-aE_Z-P3eT4u-Z8mTWITtarexIcc5GodYgUD4Jct1KbLApoNyD5Uk5J_Jmk8LapgfDwWz9m51_s5VrAM3Ov7kpubeHC-Z-TcNj6iC8AO8OgM3Ornyyowv5kRMCtMK2cF_2HBVvfwIlk4v10dEQUvkmM8TwxCr_ALH-ol4</recordid><startdate>20030202</startdate><enddate>20030202</enddate><creator>Narongwanichgarn, Wonganun</creator><creator>Misawa, Naoaki</creator><creator>Jin, Jing Hua</creator><creator>Amoako, Kingsley Kwaku</creator><creator>Kawaguchi, Emi</creator><creator>Shinjo, Toshiharu</creator><creator>Haga, Takeshi</creator><creator>Goto, Yoshitaka</creator><general>Elsevier B.V</general><general>Elsevier Science</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>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20030202</creationdate><title>Specific detection and differentiation of two subspecies of Fusobacterium necrophorum by PCR</title><author>Narongwanichgarn, Wonganun ; Misawa, Naoaki ; Jin, Jing Hua ; Amoako, Kingsley Kwaku ; Kawaguchi, Emi ; Shinjo, Toshiharu ; Haga, Takeshi ; Goto, Yoshitaka</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c488t-1472cf1bc58bf2a1923e20b0fd78264e99f2eb0c0a4d751783e597f9a50c62f73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Bacteriological methods and techniques used in bacteriology</topic><topic>Bacteriology</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cloning, Molecular</topic><topic>Cluster Analysis</topic><topic>DNA Primers - chemistry</topic><topic>DNA Primers - genetics</topic><topic>DNA, Bacterial - chemistry</topic><topic>DNA, Bacterial - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fusobacterium necrophorum</topic><topic>Fusobacterium necrophorum - chemistry</topic><topic>Fusobacterium necrophorum - classification</topic><topic>Fusobacterium necrophorum - genetics</topic><topic>Microbiology</topic><topic>Molecular Sequence Data</topic><topic>Nucleic Acid Hybridization</topic><topic>PCR</topic><topic>Phylogeny</topic><topic>Polymerase Chain Reaction - veterinary</topic><topic>Random Amplified Polymorphic DNA Technique - veterinary</topic><topic>Sequence Analysis, DNA</topic><topic>Specific detection and differentiation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Narongwanichgarn, Wonganun</creatorcontrib><creatorcontrib>Misawa, Naoaki</creatorcontrib><creatorcontrib>Jin, Jing Hua</creatorcontrib><creatorcontrib>Amoako, Kingsley Kwaku</creatorcontrib><creatorcontrib>Kawaguchi, Emi</creatorcontrib><creatorcontrib>Shinjo, Toshiharu</creatorcontrib><creatorcontrib>Haga, Takeshi</creatorcontrib><creatorcontrib>Goto, Yoshitaka</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>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>Veterinary microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Narongwanichgarn, Wonganun</au><au>Misawa, Naoaki</au><au>Jin, Jing Hua</au><au>Amoako, Kingsley Kwaku</au><au>Kawaguchi, Emi</au><au>Shinjo, Toshiharu</au><au>Haga, Takeshi</au><au>Goto, Yoshitaka</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Specific detection and differentiation of two subspecies of Fusobacterium necrophorum by PCR</atitle><jtitle>Veterinary microbiology</jtitle><addtitle>Vet Microbiol</addtitle><date>2003-02-02</date><risdate>2003</risdate><volume>91</volume><issue>2</issue><spage>183</spage><epage>195</epage><pages>183-195</pages><issn>0378-1135</issn><eissn>1873-2542</eissn><coden>VMICDQ</coden><abstract>The phylogenic relationships of two subspecies of
Fusobacterium necrophorum were investigated by randomly amplified polymorphism DNA–polymerase chain reaction (RAPD–PCR). With each of the 12 random primers, the DNA fingerprints generated were subjected to cluster analysis for dendrograms. The analysis indicated that twelve strains were organized into two major clusters, and that all strains of each subspecies were confined to one cluster. Furthermore, two of the random primers examined each generated a unique band in
F. n. necrophorum strains. We cloned these specific bands and determined the nucleotide sequences. A search for amino acid sequence homologies revealed that the two specific fragments had significant homology to the
rpoB gene of
Lactococcus lactis subsp.
lactis and the hemagglutinin-related protein gene of
Ralstonia solanacearum, respectively. New specific primers designed for the
rpoB gene were able to amplify 900
bp fragments from both subspecies. However, the specific primers designed for the hemagglutinin-related protein gene amplified only a 250
bp fragment of the genome of the
F. n. necrophorum strains, suggesting that this gene is unique to
F. n. necrophorum. These results were further confirmed by dot blot hybridization. Finally, a one-step duplex PCR technique in a single tube for the rapid detection and differentiation of the
F. necrophorum subspecies was developed.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>12458167</pmid><doi>10.1016/S0378-1135(02)00295-X</doi><tpages>13</tpages></addata></record> |
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| identifier | ISSN: 0378-1135 |
| ispartof | Veterinary microbiology, 2003-02, Vol.91 (2), p.183-195 |
| issn | 0378-1135 1873-2542 |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Bacteriological methods and techniques used in bacteriology Bacteriology Base Sequence Biological and medical sciences Cloning, Molecular Cluster Analysis DNA Primers - chemistry DNA Primers - genetics DNA, Bacterial - chemistry DNA, Bacterial - genetics Fundamental and applied biological sciences. Psychology Fusobacterium necrophorum Fusobacterium necrophorum - chemistry Fusobacterium necrophorum - classification Fusobacterium necrophorum - genetics Microbiology Molecular Sequence Data Nucleic Acid Hybridization PCR Phylogeny Polymerase Chain Reaction - veterinary Random Amplified Polymorphic DNA Technique - veterinary Sequence Analysis, DNA Specific detection and differentiation |
| title | Specific detection and differentiation of two subspecies of Fusobacterium necrophorum by PCR |
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