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Detection and quantitation of two cucurbit criniviruses in mixed infection by real-time RT-PCR

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dc.contributor.author Abrahamian P.E.
dc.contributor.author Seblani R.
dc.contributor.author Sobh H.
dc.contributor.author Abou-Jawdah Y.
dc.contributor.editor
dc.date 2013
dc.date.accessioned 2017-10-18T13:13:44Z
dc.date.available 2017-10-18T13:13:44Z
dc.date.issued 2013
dc.identifier 10.1016/j.jviromet.2013.06.004
dc.identifier.issn 1660934
dc.identifier.uri http://hdl.handle.net/10938/20177
dc.description.abstract Cucurbit chlorotic yellows virus (CCYV) and Cucurbit yellow stunting disorder virus (CYSDV) are whitefly-transmitted criniviruses infecting cucurbit crops inducing similar symptoms. Single and multiplex RT-PCR protocols were developed and evaluated on cucurbit samples collected from commercial greenhouses. Primers and probes were designed from the highly conserved heat shock protein 70 homolog (Hsp70h) gene. Conventional RT-PCR and multiplex RT-PCR assays showed high specificity and suitability for routine screening. TaqMan-based quantitative real-time RT-PCR (RT-qPCR) protocols were also developed for the detection and quantitation of both viruses occurring in single or mixed infection. The assays proved to be highly specific with no cross amplification. RT-qPCR assays showed a 100-1000 times improved sensitivity over conventional RT-PCR. Virus titers in mixed infections were compared to singly infected plants by RT-qPCR. CYSDV and CCYV titers decreased in double infected plants. This paper reports highly specific conventional RT-PCR and quantitative real-time PCR assays for detection, quantitation and differentiation between two closely related cucurbit-infecting criniviruses. © 2013 Elsevier B.V.
dc.format.extent
dc.format.extent Pages: (320-326)
dc.language English
dc.publisher AMSTERDAM
dc.relation.ispartof Publication Name: Journal of Virological Methods; Publication Year: 2013; Volume: 193; no. 2; Pages: (320-326);
dc.source Scopus
dc.title Detection and quantitation of two cucurbit criniviruses in mixed infection by real-time RT-PCR
dc.type Article
dc.contributor.affiliation Abrahamian, P.E., Department of Agricultural Sciences, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut 1107 2020, Lebanon
dc.contributor.affiliation Seblani, R., Department of Agricultural Sciences, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut 1107 2020, Lebanon
dc.contributor.affiliation Sobh, H., Department of Agricultural Sciences, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut 1107 2020, Lebanon
dc.contributor.affiliation Abou-Jawdah, Y., Department of Agricultural Sciences, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut 1107 2020, Lebanon
dc.contributor.authorAddress Abrahamian, P.E.; Department of Agricultural Sciences, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut 1107 2020, Lebanon; email: peterabrahamian@gmail.com
dc.contributor.authorCorporate University: American University of Beirut; Faculty: Faculty of Agricultural and Food Sciences; Department: Agricultural Sciences;
dc.contributor.authorDepartment Agricultural Sciences
dc.contributor.authorDivision
dc.contributor.authorEmail peterabrahamian@gmail.com; rewaseblani@gmail.com; hs05@aub.edu.lb; abujawyf@aub.edu.lb
dc.contributor.authorFaculty Faculty of Agricultural and Food Sciences
dc.contributor.authorInitials Abrahamian, PE
dc.contributor.authorInitials Seblani, R
dc.contributor.authorInitials Sobh, H
dc.contributor.authorInitials Abou-Jawdah, Y
dc.contributor.authorOrcidID
dc.contributor.authorReprintAddress Abou-Jawdah, Y (reprint author), Amer Univ Beirut, Fac Agr and Food Sci, Dept Agr Sci, Beirut 11072020, Lebanon.
dc.contributor.authorResearcherID
dc.contributor.authorUniversity American University of Beirut
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dc.description.citedCount 1
dc.description.citedTotWOSCount 2
dc.description.citedWOSCount 1
dc.format.extentCount 7
dc.identifier.articleNo
dc.identifier.coden JVMED
dc.identifier.pubmedID 23810855
dc.identifier.scopusID 84880679652
dc.publisher.address PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
dc.relation.ispartofFundingAgency USAID, United States Agency for International Development
dc.relation.ispartOfISOAbbr J. Virol. Methods
dc.relation.ispartOfIssue 2
dc.relation.ispartofPubTitle Journal of Virological Methods
dc.relation.ispartofPubTitleAbbr J. Virol. Methods
dc.relation.ispartOfVolume 193
dc.source.ID WOS:000325190400010
dc.type.publication Journal
dc.subject.otherAuthKeyword Ccyv
dc.subject.otherAuthKeyword cucurbit chlorotic yellows virus
dc.subject.otherAuthKeyword cucurbit yellow stunting disorder virus
dc.subject.otherAuthKeyword cysdv
dc.subject.otherAuthKeyword multiplex detection
dc.subject.otherAuthKeyword rt-qpcr
dc.subject.otherChemCAS Dna primers
dc.subject.otherChemCAS hsp70 heat-shock proteins
dc.subject.otherChemCAS oligonucleotide probes
dc.subject.otherChemCAS viral proteins
dc.subject.otherIndex Heat shock protein 70
dc.subject.otherIndex article
dc.subject.otherIndex controlled study
dc.subject.otherIndex crinivirus
dc.subject.otherIndex cucurbit chlorotic yellows virus
dc.subject.otherIndex cucurbitaceae
dc.subject.otherIndex gene amplification
dc.subject.otherIndex hsp70h gene
dc.subject.otherIndex intermethod comparison
dc.subject.otherIndex mixed infection
dc.subject.otherIndex molecular probe
dc.subject.otherIndex nonhuman
dc.subject.otherIndex priority journal
dc.subject.otherIndex reverse transcription polymerase chain reaction
dc.subject.otherIndex screening test
dc.subject.otherIndex sensitivity and specificity
dc.subject.otherIndex virus gene
dc.subject.otherIndex virus identification
dc.subject.otherIndex ccyv
dc.subject.otherIndex cucurbit chlorotic yellows virus
dc.subject.otherIndex cucurbit yellow stunting disorder virus
dc.subject.otherIndex cysdv
dc.subject.otherIndex multiplex detection
dc.subject.otherIndex rt-qpcr
dc.subject.otherIndex coinfection
dc.subject.otherIndex crinivirus
dc.subject.otherIndex cucurbita
dc.subject.otherIndex dna primers
dc.subject.otherIndex hsp70 heat-shock proteins
dc.subject.otherIndex multiplex polymerase chain reaction
dc.subject.otherIndex oligonucleotide probes
dc.subject.otherIndex plant diseases
dc.subject.otherIndex real-time polymerase chain reaction
dc.subject.otherIndex reverse transcriptase polymerase chain reaction
dc.subject.otherIndex sensitivity and specificity
dc.subject.otherIndex viral proteins
dc.subject.otherKeywordPlus Stunting-disorder-virus
dc.subject.otherKeywordPlus chlorotic-yellows-virus
dc.subject.otherKeywordPlus bemisia-tabaci
dc.subject.otherKeywordPlus 1st report
dc.subject.otherKeywordPlus host-range
dc.subject.otherKeywordPlus cucumber
dc.subject.otherKeywordPlus crops
dc.subject.otherKeywordPlus melon
dc.subject.otherKeywordPlus closteroviruses
dc.subject.otherKeywordPlus coinfection
dc.subject.otherWOS Biochemical research methods
dc.subject.otherWOS biotechnology and applied microbiology
dc.subject.otherWOS virology
dc.identifier.doi http://dx.doi.org/10.1016/j.jviromet.2013.06.004


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