Loading…

Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control

Two multiplex RT-PCR assays with specific coamplification of plant mRNA as an internal control from total nucleic acids are described for the parallel detection of Apple chlorotic leaf spot virus, Apple stem pitting virus, Apple mosaic virus and Apple stem grooving virus. All are important economica...

Full description

Saved in:
Bibliographic Details
Published in:Journal of virological methods 2002, Vol.99 (1), p.81-92
Main Authors: Menzel, W., Jelkmann, W., Maiss, E.
Format: Article
Language:English
Subjects:
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-c474t-7eca5b9583b6144d9a4165c058fbbc2b5cb59c94f6a6bbdae70df0b015c8936d3
cites cdi_FETCH-LOGICAL-c474t-7eca5b9583b6144d9a4165c058fbbc2b5cb59c94f6a6bbdae70df0b015c8936d3
container_end_page 92
container_issue 1
container_start_page 81
container_title Journal of virological methods
container_volume 99
creator Menzel, W.
Jelkmann, W.
Maiss, E.
description Two multiplex RT-PCR assays with specific coamplification of plant mRNA as an internal control from total nucleic acids are described for the parallel detection of Apple chlorotic leaf spot virus, Apple stem pitting virus, Apple mosaic virus and Apple stem grooving virus. All are important economically and common pathogens in commercial apple and pear cultivars, except for Apple mosaic virus. Four virus specific primer pairs and one primer pair which allows the specific amplification of mRNA of the mitochondrial nad5 gene are described. Specificity of all primer pairs was confirmed by sequencing the RT-PCR products. A range of different virus isolates from various geographic origins could be detected by these multiplex RT-PCR assays all year round. Viruses were detected reliably in composite extracts at a ratio of one part total nucleic acid extract from an infected sample mixed with 39 parts of extract from healthy samples. The use of the internal control minimizes the risk of obtaining false negative RT-PCR results, which is desirable for routine testing, and avoids the need to eliminate contaminating DNA in extracts. To our knowledge, this is the first report on the use of a specific internal RNA control from total nucleic acids. The multiplex RT-PCR assays described are reliable, rapid and sensitive methods for the detection of these viruses, and may replace techniques need commonly like indexing by woody indicators or ELISA.
doi_str_mv 10.1016/S0166-0934(01)00381-0
format article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_71338349</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0166093401003810</els_id><sourcerecordid>71338349</sourcerecordid><originalsourceid>FETCH-LOGICAL-c474t-7eca5b9583b6144d9a4165c058fbbc2b5cb59c94f6a6bbdae70df0b015c8936d3</originalsourceid><addsrcrecordid>eNqF0U1vFSEUBmBiNPa2-hM0bDS6mHooH8OsTHOtH0lTzbWuCTAQMcwwhZna---lvbd22Q0k5DmHk_Mi9IrAMQEiPvysh2igo-wdkPcAVJIGnqAVkW1XnyV7ilb_yQE6LOUPAPCW0ufogBAhGQWxQlef3OzsHNKIk8c-LRnraYoOX4e8FFew2eJhiXOobzd4c9n8WG-wLkVvC_4b5t_YJj1MMfhg9X2XKepxxsPm4rRKHMbZ5VHHKsc5p_gCPfM6Fvdyfx-hX5_PLtdfm_PvX76tT88by1o2N62zmpuOS2oEYazvNCOCW-DSG2NPDLeGd7ZjXmhhTK9dC70HA4Rb2VHR0yP0dtd3yulqcWVWQyjWxTqcS0tRLaFUUtY9CokkHTDWVsh30OZUSnZeTTkMOm8VAXUbiroLRd1uXAFRd6EoqHWv9x8sZnD9Q9U-hQre7IEuVkef9WhDeXCUAUh5Ut3HnXN1b9fBZVVscKN1fcg1RNWn8Mgo_wAdpKnf</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>18190447</pqid></control><display><type>article</type><title>Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control</title><source>ScienceDirect Freedom Collection 2022-2024</source><creator>Menzel, W. ; Jelkmann, W. ; Maiss, E.</creator><creatorcontrib>Menzel, W. ; Jelkmann, W. ; Maiss, E.</creatorcontrib><description>Two multiplex RT-PCR assays with specific coamplification of plant mRNA as an internal control from total nucleic acids are described for the parallel detection of Apple chlorotic leaf spot virus, Apple stem pitting virus, Apple mosaic virus and Apple stem grooving virus. All are important economically and common pathogens in commercial apple and pear cultivars, except for Apple mosaic virus. Four virus specific primer pairs and one primer pair which allows the specific amplification of mRNA of the mitochondrial nad5 gene are described. Specificity of all primer pairs was confirmed by sequencing the RT-PCR products. A range of different virus isolates from various geographic origins could be detected by these multiplex RT-PCR assays all year round. Viruses were detected reliably in composite extracts at a ratio of one part total nucleic acid extract from an infected sample mixed with 39 parts of extract from healthy samples. The use of the internal control minimizes the risk of obtaining false negative RT-PCR results, which is desirable for routine testing, and avoids the need to eliminate contaminating DNA in extracts. To our knowledge, this is the first report on the use of a specific internal RNA control from total nucleic acids. The multiplex RT-PCR assays described are reliable, rapid and sensitive methods for the detection of these viruses, and may replace techniques need commonly like indexing by woody indicators or ELISA.</description><identifier>ISSN: 0166-0934</identifier><identifier>EISSN: 1879-0984</identifier><identifier>DOI: 10.1016/S0166-0934(01)00381-0</identifier><identifier>PMID: 11684306</identifier><identifier>CODEN: JVMEDH</identifier><language>eng</language><publisher>London: Elsevier B.V</publisher><subject>Apple chlorotic leaf spot virus ; Apple mosaic virus ; Apple stem grooving virus ; Apple stem pitting virus ; Apple virus detection ; Biological and medical sciences ; Fundamental and applied biological sciences. Psychology ; Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control ; Internal control ; Malus - genetics ; Malus - virology ; Microbiology ; Multiplex RT-PCR ; nad5 gene ; Phytopathology. Animal pests. Plant and forest protection ; Plant Diseases - virology ; Plant Viruses - genetics ; Plant Viruses - isolation &amp; purification ; Plant viruses and viroids ; Reference Standards ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger - genetics ; RNA, Plant - genetics ; RNA, Viral - analysis ; Sensitivity and Specificity ; Techniques used in virology ; Virology</subject><ispartof>Journal of virological methods, 2002, Vol.99 (1), p.81-92</ispartof><rights>2002 Elsevier Science B.V.</rights><rights>2002 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c474t-7eca5b9583b6144d9a4165c058fbbc2b5cb59c94f6a6bbdae70df0b015c8936d3</citedby><cites>FETCH-LOGICAL-c474t-7eca5b9583b6144d9a4165c058fbbc2b5cb59c94f6a6bbdae70df0b015c8936d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,4024,27923,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=13400882$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11684306$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Menzel, W.</creatorcontrib><creatorcontrib>Jelkmann, W.</creatorcontrib><creatorcontrib>Maiss, E.</creatorcontrib><title>Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control</title><title>Journal of virological methods</title><addtitle>J Virol Methods</addtitle><description>Two multiplex RT-PCR assays with specific coamplification of plant mRNA as an internal control from total nucleic acids are described for the parallel detection of Apple chlorotic leaf spot virus, Apple stem pitting virus, Apple mosaic virus and Apple stem grooving virus. All are important economically and common pathogens in commercial apple and pear cultivars, except for Apple mosaic virus. Four virus specific primer pairs and one primer pair which allows the specific amplification of mRNA of the mitochondrial nad5 gene are described. Specificity of all primer pairs was confirmed by sequencing the RT-PCR products. A range of different virus isolates from various geographic origins could be detected by these multiplex RT-PCR assays all year round. Viruses were detected reliably in composite extracts at a ratio of one part total nucleic acid extract from an infected sample mixed with 39 parts of extract from healthy samples. The use of the internal control minimizes the risk of obtaining false negative RT-PCR results, which is desirable for routine testing, and avoids the need to eliminate contaminating DNA in extracts. To our knowledge, this is the first report on the use of a specific internal RNA control from total nucleic acids. The multiplex RT-PCR assays described are reliable, rapid and sensitive methods for the detection of these viruses, and may replace techniques need commonly like indexing by woody indicators or ELISA.</description><subject>Apple chlorotic leaf spot virus</subject><subject>Apple mosaic virus</subject><subject>Apple stem grooving virus</subject><subject>Apple stem pitting virus</subject><subject>Apple virus detection</subject><subject>Biological and medical sciences</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control</subject><subject>Internal control</subject><subject>Malus - genetics</subject><subject>Malus - virology</subject><subject>Microbiology</subject><subject>Multiplex RT-PCR</subject><subject>nad5 gene</subject><subject>Phytopathology. Animal pests. Plant and forest protection</subject><subject>Plant Diseases - virology</subject><subject>Plant Viruses - genetics</subject><subject>Plant Viruses - isolation &amp; purification</subject><subject>Plant viruses and viroids</subject><subject>Reference Standards</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Plant - genetics</subject><subject>RNA, Viral - analysis</subject><subject>Sensitivity and Specificity</subject><subject>Techniques used in virology</subject><subject>Virology</subject><issn>0166-0934</issn><issn>1879-0984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><recordid>eNqF0U1vFSEUBmBiNPa2-hM0bDS6mHooH8OsTHOtH0lTzbWuCTAQMcwwhZna---lvbd22Q0k5DmHk_Mi9IrAMQEiPvysh2igo-wdkPcAVJIGnqAVkW1XnyV7ilb_yQE6LOUPAPCW0ufogBAhGQWxQlef3OzsHNKIk8c-LRnraYoOX4e8FFew2eJhiXOobzd4c9n8WG-wLkVvC_4b5t_YJj1MMfhg9X2XKepxxsPm4rRKHMbZ5VHHKsc5p_gCPfM6Fvdyfx-hX5_PLtdfm_PvX76tT88by1o2N62zmpuOS2oEYazvNCOCW-DSG2NPDLeGd7ZjXmhhTK9dC70HA4Rb2VHR0yP0dtd3yulqcWVWQyjWxTqcS0tRLaFUUtY9CokkHTDWVsh30OZUSnZeTTkMOm8VAXUbiroLRd1uXAFRd6EoqHWv9x8sZnD9Q9U-hQre7IEuVkef9WhDeXCUAUh5Ut3HnXN1b9fBZVVscKN1fcg1RNWn8Mgo_wAdpKnf</recordid><startdate>2002</startdate><enddate>2002</enddate><creator>Menzel, W.</creator><creator>Jelkmann, W.</creator><creator>Maiss, E.</creator><general>Elsevier B.V</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>7QO</scope><scope>7T7</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>2002</creationdate><title>Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control</title><author>Menzel, W. ; Jelkmann, W. ; Maiss, E.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c474t-7eca5b9583b6144d9a4165c058fbbc2b5cb59c94f6a6bbdae70df0b015c8936d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Apple chlorotic leaf spot virus</topic><topic>Apple mosaic virus</topic><topic>Apple stem grooving virus</topic><topic>Apple stem pitting virus</topic><topic>Apple virus detection</topic><topic>Biological and medical sciences</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control</topic><topic>Internal control</topic><topic>Malus - genetics</topic><topic>Malus - virology</topic><topic>Microbiology</topic><topic>Multiplex RT-PCR</topic><topic>nad5 gene</topic><topic>Phytopathology. Animal pests. Plant and forest protection</topic><topic>Plant Diseases - virology</topic><topic>Plant Viruses - genetics</topic><topic>Plant Viruses - isolation &amp; purification</topic><topic>Plant viruses and viroids</topic><topic>Reference Standards</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Plant - genetics</topic><topic>RNA, Viral - analysis</topic><topic>Sensitivity and Specificity</topic><topic>Techniques used in virology</topic><topic>Virology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Menzel, W.</creatorcontrib><creatorcontrib>Jelkmann, W.</creatorcontrib><creatorcontrib>Maiss, E.</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>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Menzel, W.</au><au>Jelkmann, W.</au><au>Maiss, E.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>2002</date><risdate>2002</risdate><volume>99</volume><issue>1</issue><spage>81</spage><epage>92</epage><pages>81-92</pages><issn>0166-0934</issn><eissn>1879-0984</eissn><coden>JVMEDH</coden><abstract>Two multiplex RT-PCR assays with specific coamplification of plant mRNA as an internal control from total nucleic acids are described for the parallel detection of Apple chlorotic leaf spot virus, Apple stem pitting virus, Apple mosaic virus and Apple stem grooving virus. All are important economically and common pathogens in commercial apple and pear cultivars, except for Apple mosaic virus. Four virus specific primer pairs and one primer pair which allows the specific amplification of mRNA of the mitochondrial nad5 gene are described. Specificity of all primer pairs was confirmed by sequencing the RT-PCR products. A range of different virus isolates from various geographic origins could be detected by these multiplex RT-PCR assays all year round. Viruses were detected reliably in composite extracts at a ratio of one part total nucleic acid extract from an infected sample mixed with 39 parts of extract from healthy samples. The use of the internal control minimizes the risk of obtaining false negative RT-PCR results, which is desirable for routine testing, and avoids the need to eliminate contaminating DNA in extracts. To our knowledge, this is the first report on the use of a specific internal RNA control from total nucleic acids. The multiplex RT-PCR assays described are reliable, rapid and sensitive methods for the detection of these viruses, and may replace techniques need commonly like indexing by woody indicators or ELISA.</abstract><cop>London</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>11684306</pmid><doi>10.1016/S0166-0934(01)00381-0</doi><tpages>12</tpages></addata></record>
fulltext fulltext
identifier ISSN: 0166-0934
ispartof Journal of virological methods, 2002, Vol.99 (1), p.81-92
issn 0166-0934
1879-0984
language eng
recordid cdi_proquest_miscellaneous_71338349
source ScienceDirect Freedom Collection 2022-2024
subjects Apple chlorotic leaf spot virus
Apple mosaic virus
Apple stem grooving virus
Apple stem pitting virus
Apple virus detection
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control
Internal control
Malus - genetics
Malus - virology
Microbiology
Multiplex RT-PCR
nad5 gene
Phytopathology. Animal pests. Plant and forest protection
Plant Diseases - virology
Plant Viruses - genetics
Plant Viruses - isolation & purification
Plant viruses and viroids
Reference Standards
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger - genetics
RNA, Plant - genetics
RNA, Viral - analysis
Sensitivity and Specificity
Techniques used in virology
Virology
title Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-24T20%3A02%3A06IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Detection%20of%20four%20apple%20viruses%20by%20multiplex%20RT-PCR%20assays%20with%20coamplification%20of%20plant%20mRNA%20as%20internal%20control&rft.jtitle=Journal%20of%20virological%20methods&rft.au=Menzel,%20W.&rft.date=2002&rft.volume=99&rft.issue=1&rft.spage=81&rft.epage=92&rft.pages=81-92&rft.issn=0166-0934&rft.eissn=1879-0984&rft.coden=JVMEDH&rft_id=info:doi/10.1016/S0166-0934(01)00381-0&rft_dat=%3Cproquest_cross%3E71338349%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c474t-7eca5b9583b6144d9a4165c058fbbc2b5cb59c94f6a6bbdae70df0b015c8936d3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=18190447&rft_id=info:pmid/11684306&rfr_iscdi=true