Use of Fluorescent Microsphere-Based Assay for Detection of Three Cucurbit-Infecting Viruses
In this study, we describe multiplex polymerase chain reaction (PCR) coupled with the LiquiChip assay for the identification of Zucchini yellow mosaic virus, Cucumber green mottle mosaic virus, and Cucumber mosaic virus by coamplification with plant mRNA as an internal control. Multiplex reverse-tra...
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| Veröffentlicht in: | Plant disease 2018-11, Vol.102 (11), p.2324-2329 |
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| container_title | Plant disease |
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| creator | Kuan, Cheng-Ping Chang, Wen-Shi Yang, Tso-Chi |
| description | In this study, we describe multiplex polymerase chain reaction (PCR) coupled with the LiquiChip assay for the identification of Zucchini yellow mosaic virus, Cucumber green mottle mosaic virus, and Cucumber mosaic virus by coamplification with plant mRNA as an internal control. Multiplex reverse-transcription (RT)-PCR products were subjected to allele-specific primer extension, then hybridized to carboxylated microspheres with unique fluorescent identifiers followed by detection using the LiquiChip 200 workstation. This assay is highly specific for distinguishing individual viruses from a mixed viral population and is 10 times more sensitive than multiplex RT-PCR. In addition, the establishment of this method enabled the detection of cucurbit viruses in field samples. |
| doi_str_mv | 10.1094/PDIS-09-17-1355-RE |
| format | Article |
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In addition, the establishment of this method enabled the detection of cucurbit viruses in field samples.</description><subject>Cucumovirus - genetics</subject><subject>Cucumovirus - isolation & purification</subject><subject>Cucurbita - virology</subject><subject>DNA Primers - genetics</subject><subject>Fluorescent Dyes</subject><subject>Microspheres</subject><subject>Multiplex Polymerase Chain Reaction</subject><subject>Plant Diseases - virology</subject><subject>Plant Viruses - genetics</subject><subject>Plant Viruses - isolation & purification</subject><subject>Potyvirus - genetics</subject><subject>Potyvirus - isolation & purification</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Sensitivity and Specificity</subject><subject>Tobamovirus - genetics</subject><subject>Tobamovirus - isolation & purification</subject><issn>0191-2917</issn><issn>1943-7692</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kMtOwzAQRS0EgvL4ARbISzaGGTuu4yX0AZVAIGhZIVl5jCEoTYqdLPh7GvFYzWLuudI9jJ0iXCDY5PJxungWYAUagUpr8TTbYSO0iRJmbOUuGwFaFNKiOWCHMX4AQJKM0312oEBK1KkasddVJN56Pq_7NlAsqOn4fVWENm7eKZC4ziKV_CrG7Iv7NvApdVR0VdsM0PI9EPFJX_QhrzqxaPzwa974SxX6SPGY7fmsjnTye4_Yaj5bTm7F3cPNYnJ1JwqVmE54bwvEUpkSDOaUFuQJMkl2nGvw6FWic0i11Eqh8mkGpbGgdVpqzAzJXB2x85_eTWg_e4qdW1fbKXWdNdT20UmEVGoNaLZR-RMdJsZA3m1Ctc7Cl0Nwg1U3WHVgHRo3WHVPsy109tvf52sq_5E_jeobWg1zdQ</recordid><startdate>201811</startdate><enddate>201811</enddate><creator>Kuan, Cheng-Ping</creator><creator>Chang, Wen-Shi</creator><creator>Yang, Tso-Chi</creator><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>7X8</scope><orcidid>https://orcid.org/0000-0002-8970-7111</orcidid></search><sort><creationdate>201811</creationdate><title>Use of Fluorescent Microsphere-Based Assay for Detection of Three Cucurbit-Infecting Viruses</title><author>Kuan, Cheng-Ping ; Chang, Wen-Shi ; Yang, Tso-Chi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c347t-ff9c11d37d071be8cefe0a2e96b50f1f345b085253313f8a0d790558d51a7e2b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Cucumovirus - genetics</topic><topic>Cucumovirus - isolation & purification</topic><topic>Cucurbita - virology</topic><topic>DNA Primers - genetics</topic><topic>Fluorescent Dyes</topic><topic>Microspheres</topic><topic>Multiplex Polymerase Chain Reaction</topic><topic>Plant Diseases - virology</topic><topic>Plant Viruses - genetics</topic><topic>Plant Viruses - isolation & purification</topic><topic>Potyvirus - genetics</topic><topic>Potyvirus - isolation & purification</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Sensitivity and Specificity</topic><topic>Tobamovirus - genetics</topic><topic>Tobamovirus - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kuan, Cheng-Ping</creatorcontrib><creatorcontrib>Chang, Wen-Shi</creatorcontrib><creatorcontrib>Yang, Tso-Chi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Plant disease</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kuan, Cheng-Ping</au><au>Chang, Wen-Shi</au><au>Yang, Tso-Chi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Use of Fluorescent Microsphere-Based Assay for Detection of Three Cucurbit-Infecting Viruses</atitle><jtitle>Plant disease</jtitle><addtitle>Plant Dis</addtitle><date>2018-11</date><risdate>2018</risdate><volume>102</volume><issue>11</issue><spage>2324</spage><epage>2329</epage><pages>2324-2329</pages><issn>0191-2917</issn><eissn>1943-7692</eissn><abstract>In this study, we describe multiplex polymerase chain reaction (PCR) coupled with the LiquiChip assay for the identification of Zucchini yellow mosaic virus, Cucumber green mottle mosaic virus, and Cucumber mosaic virus by coamplification with plant mRNA as an internal control. 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| fulltext | fulltext |
| identifier | ISSN: 0191-2917 |
| ispartof | Plant disease, 2018-11, Vol.102 (11), p.2324-2329 |
| issn | 0191-2917 1943-7692 |
| language | eng |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection; American Phytopathological Society Journal Back Issues |
| subjects | Cucumovirus - genetics Cucumovirus - isolation & purification Cucurbita - virology DNA Primers - genetics Fluorescent Dyes Microspheres Multiplex Polymerase Chain Reaction Plant Diseases - virology Plant Viruses - genetics Plant Viruses - isolation & purification Potyvirus - genetics Potyvirus - isolation & purification Reverse Transcriptase Polymerase Chain Reaction Sensitivity and Specificity Tobamovirus - genetics Tobamovirus - isolation & purification |
| title | Use of Fluorescent Microsphere-Based Assay for Detection of Three Cucurbit-Infecting Viruses |
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