Detection of Botrytis cinerea by loop‐mediated isothermal amplification
Aims: To develop a sensitive, rapid and simple method for detection of Botrytis cinerea based on loop‐mediated isothermal amplification (LAMP) that would be suitable for use outside a conventional laboratory setting. Methods and Results: A LAMP assay was designed based on the intergenic spacer of th...
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| Veröffentlicht in: | Letters in applied microbiology 2010-12, Vol.51 (6), p.650-657 |
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| creator | Tomlinson, J.A Dickinson, M.J Boonham, N |
| description | Aims: To develop a sensitive, rapid and simple method for detection of Botrytis cinerea based on loop‐mediated isothermal amplification (LAMP) that would be suitable for use outside a conventional laboratory setting. Methods and Results: A LAMP assay was designed based on the intergenic spacer of the B. cinerea nuclear ribosomal DNA (rDNA). The resulting assay was characterized in terms of sensitivity and specificity using DNA extracted from cultures. The assay consistently amplified 65 pg B. cinerea DNA. No cross‐reactivity was observed with a range of other fungal pathogens, with the exception of the closely related species Botrytis pelargonii. Use of a novel real‐time LAMP platform (the OptiGene Genie I) allowed detection of B. cinerea in infected rose petals, with amplification occurring in |
| doi_str_mv | 10.1111/j.1472-765X.2010.02949.x |
| format | Article |
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Methods and Results: A LAMP assay was designed based on the intergenic spacer of the B. cinerea nuclear ribosomal DNA (rDNA). The resulting assay was characterized in terms of sensitivity and specificity using DNA extracted from cultures. The assay consistently amplified 65 pg B. cinerea DNA. No cross‐reactivity was observed with a range of other fungal pathogens, with the exception of the closely related species Botrytis pelargonii. Use of a novel real‐time LAMP platform (the OptiGene Genie I) allowed detection of B. cinerea in infected rose petals, with amplification occurring in <15 min. Conclusions: The LAMP assay that was developed is suitable for rapid detection of B. cinerea in infected plant material. Significance and Impact of the Study: The LAMP method combines the sensitivity and specificity of nucleic acid‐based methods with simplified equipment and a reduced reaction time. These features make the method potentially suitable for on‐site use, where the results of testing could help to inform decisions regarding the storage and processing of commodities affected by B. cinerea, such as cut flowers, fruit and vegetables.</description><identifier>ISSN: 0266-8254</identifier><identifier>EISSN: 1472-765X</identifier><identifier>DOI: 10.1111/j.1472-765X.2010.02949.x</identifier><identifier>PMID: 21029140</identifier><identifier>CODEN: LAMIE7</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Biological and medical sciences ; Botrytis ; Botrytis - isolation & purification ; Botrytis cinerea ; corolla ; cut flowers ; DNA Primers ; DNA, Fungal - isolation & purification ; DNA, Ribosomal - isolation & purification ; Fundamental and applied biological sciences. Psychology ; fungi ; grey mould ; intergenic DNA ; isothermal amplification ; Microbiology ; Nucleic Acid Amplification Techniques - methods ; pathogens ; Plant Diseases - microbiology ; quantitative detection ; rapid methods ; ribosomal DNA ; Rosa - microbiology ; Sensitivity and Specificity ; vegetables</subject><ispartof>Letters in applied microbiology, 2010-12, Vol.51 (6), p.650-657</ispartof><rights>2010 British Crown Copyright. Letters in Applied Microbiology 51, 650–657 © 2010 The Society for Applied Microbiology</rights><rights>2015 INIST-CNRS</rights><rights>2010 British Crown Copyright. Letters in Applied Microbiology 51, 650-657 © 2010 The Society for Applied Microbiology.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4299-85d42044aa153544016ff2458c79099571e372ecb0f826928c6eaf1f160ef19b3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1472-765X.2010.02949.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1472-765X.2010.02949.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=23424182$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21029140$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tomlinson, J.A</creatorcontrib><creatorcontrib>Dickinson, M.J</creatorcontrib><creatorcontrib>Boonham, N</creatorcontrib><title>Detection of Botrytis cinerea by loop‐mediated isothermal amplification</title><title>Letters in applied microbiology</title><addtitle>Lett Appl Microbiol</addtitle><description>Aims: To develop a sensitive, rapid and simple method for detection of Botrytis cinerea based on loop‐mediated isothermal amplification (LAMP) that would be suitable for use outside a conventional laboratory setting. Methods and Results: A LAMP assay was designed based on the intergenic spacer of the B. cinerea nuclear ribosomal DNA (rDNA). The resulting assay was characterized in terms of sensitivity and specificity using DNA extracted from cultures. The assay consistently amplified 65 pg B. cinerea DNA. No cross‐reactivity was observed with a range of other fungal pathogens, with the exception of the closely related species Botrytis pelargonii. Use of a novel real‐time LAMP platform (the OptiGene Genie I) allowed detection of B. cinerea in infected rose petals, with amplification occurring in <15 min. Conclusions: The LAMP assay that was developed is suitable for rapid detection of B. cinerea in infected plant material. Significance and Impact of the Study: The LAMP method combines the sensitivity and specificity of nucleic acid‐based methods with simplified equipment and a reduced reaction time. These features make the method potentially suitable for on‐site use, where the results of testing could help to inform decisions regarding the storage and processing of commodities affected by B. cinerea, such as cut flowers, fruit and vegetables.</description><subject>Biological and medical sciences</subject><subject>Botrytis</subject><subject>Botrytis - isolation & purification</subject><subject>Botrytis cinerea</subject><subject>corolla</subject><subject>cut flowers</subject><subject>DNA Primers</subject><subject>DNA, Fungal - isolation & purification</subject><subject>DNA, Ribosomal - isolation & purification</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>fungi</subject><subject>grey mould</subject><subject>intergenic DNA</subject><subject>isothermal amplification</subject><subject>Microbiology</subject><subject>Nucleic Acid Amplification Techniques - methods</subject><subject>pathogens</subject><subject>Plant Diseases - microbiology</subject><subject>quantitative detection</subject><subject>rapid methods</subject><subject>ribosomal DNA</subject><subject>Rosa - microbiology</subject><subject>Sensitivity and Specificity</subject><subject>vegetables</subject><issn>0266-8254</issn><issn>1472-765X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkctOGzEUhi1UBCHtK8BsEKsJvhzb4wULbi1IqbpoI3VnOY4NjmbiMJ4IsuMR-ow8ST0kwKqqN7Z8vv_48iFUEDwieZzORwQkLaXgv0cU511MFajR0w4avBc-oQGmQpQV5bCPDlKaY4wrQtUe2qckBwjgAbq9cp2zXYiLIvriInbtugupsGHhWmeK6bqoY1y-PP9p3CyYzs2KkGJ379rG1IVplnXwwZo-_xntelMn92U7D9Hk6_Wvy5ty_OPb7eX5uLRAlSorPgOKAYwhnHEATIT3FHhlpcJKcUkck9TZKfYVFYpWVjjjiScCO0_UlA3Ryabvso0PK5c63YRkXV2bhYurpCsBinNF5H9JKYADY1hk8nBLrqb5oXrZhsa0a_32Txk43gImWVP71ixsSB8cAwqkopk723CPoXbr9zrBuvem57rXo3s9uvemX73pJz0-_96vcv5ok_cmanPX5jMmPzMJ2Z3kQvB_EgyTfFMuGfsLiNyc8g</recordid><startdate>201012</startdate><enddate>201012</enddate><creator>Tomlinson, J.A</creator><creator>Dickinson, M.J</creator><creator>Boonham, N</creator><general>Blackwell Publishing Ltd</general><general>Blackwell</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope></search><sort><creationdate>201012</creationdate><title>Detection of Botrytis cinerea by loop‐mediated isothermal amplification</title><author>Tomlinson, J.A ; Dickinson, M.J ; Boonham, N</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4299-85d42044aa153544016ff2458c79099571e372ecb0f826928c6eaf1f160ef19b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Biological and medical sciences</topic><topic>Botrytis</topic><topic>Botrytis - isolation & purification</topic><topic>Botrytis cinerea</topic><topic>corolla</topic><topic>cut flowers</topic><topic>DNA Primers</topic><topic>DNA, Fungal - isolation & purification</topic><topic>DNA, Ribosomal - isolation & purification</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>fungi</topic><topic>grey mould</topic><topic>intergenic DNA</topic><topic>isothermal amplification</topic><topic>Microbiology</topic><topic>Nucleic Acid Amplification Techniques - methods</topic><topic>pathogens</topic><topic>Plant Diseases - microbiology</topic><topic>quantitative detection</topic><topic>rapid methods</topic><topic>ribosomal DNA</topic><topic>Rosa - microbiology</topic><topic>Sensitivity and Specificity</topic><topic>vegetables</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Tomlinson, J.A</creatorcontrib><creatorcontrib>Dickinson, M.J</creatorcontrib><creatorcontrib>Boonham, N</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</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>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Letters in applied microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tomlinson, J.A</au><au>Dickinson, M.J</au><au>Boonham, N</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of Botrytis cinerea by loop‐mediated isothermal amplification</atitle><jtitle>Letters in applied microbiology</jtitle><addtitle>Lett Appl Microbiol</addtitle><date>2010-12</date><risdate>2010</risdate><volume>51</volume><issue>6</issue><spage>650</spage><epage>657</epage><pages>650-657</pages><issn>0266-8254</issn><eissn>1472-765X</eissn><coden>LAMIE7</coden><abstract>Aims: To develop a sensitive, rapid and simple method for detection of Botrytis cinerea based on loop‐mediated isothermal amplification (LAMP) that would be suitable for use outside a conventional laboratory setting. Methods and Results: A LAMP assay was designed based on the intergenic spacer of the B. cinerea nuclear ribosomal DNA (rDNA). The resulting assay was characterized in terms of sensitivity and specificity using DNA extracted from cultures. The assay consistently amplified 65 pg B. cinerea DNA. No cross‐reactivity was observed with a range of other fungal pathogens, with the exception of the closely related species Botrytis pelargonii. Use of a novel real‐time LAMP platform (the OptiGene Genie I) allowed detection of B. cinerea in infected rose petals, with amplification occurring in <15 min. Conclusions: The LAMP assay that was developed is suitable for rapid detection of B. cinerea in infected plant material. Significance and Impact of the Study: The LAMP method combines the sensitivity and specificity of nucleic acid‐based methods with simplified equipment and a reduced reaction time. These features make the method potentially suitable for on‐site use, where the results of testing could help to inform decisions regarding the storage and processing of commodities affected by B. cinerea, such as cut flowers, fruit and vegetables.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>21029140</pmid><doi>10.1111/j.1472-765X.2010.02949.x</doi><tpages>8</tpages></addata></record> |
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| source | Oxford University Press Journals All Titles (1996-Current); MEDLINE; Wiley Online Library Journals Frontfile Complete; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Biological and medical sciences Botrytis Botrytis - isolation & purification Botrytis cinerea corolla cut flowers DNA Primers DNA, Fungal - isolation & purification DNA, Ribosomal - isolation & purification Fundamental and applied biological sciences. Psychology fungi grey mould intergenic DNA isothermal amplification Microbiology Nucleic Acid Amplification Techniques - methods pathogens Plant Diseases - microbiology quantitative detection rapid methods ribosomal DNA Rosa - microbiology Sensitivity and Specificity vegetables |
| title | Detection of Botrytis cinerea by loop‐mediated isothermal amplification |
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