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High-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants

dc.contributor.authorOtti, G.
dc.contributor.authorBouvaine, S.
dc.contributor.authorKimata, B.
dc.contributor.authorMkamillo, G.
dc.contributor.authorKumar, P.L.
dc.contributor.authorTomlins, K.
dc.contributor.authorMaruthi, M.N.
dc.date.accessioned2019-12-04T11:03:49Z
dc.date.available2019-12-04T11:03:49Z
dc.date.issued2016-05
dc.identifier.citationOtti, G., Bouvaine, S., Kimata, B., Mkamillo, G., Kumar, P., Tomlins, K. & Maruthi, M.N. (2016). High throughput multiplex real time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants. Journal of applied microbiology 120(5), pp.1346-1356.
dc.identifier.issn1364-5072
dc.identifier.urihttps://hdl.handle.net/20.500.12478/1247
dc.descriptionFirst published: 11 March 2016
dc.description.abstractAims: To develop a multiplex TaqMan-based real-time PCR assay (qPCR) for the simultaneous detection and quantification of both RNA and DNA viruses affecting cassava (Manihot esculenta) in eastern Africa. Methods and Results: The diagnostic assay was developed for two RNA viruses; Cassava brown streak virus (CBSV) and Uganda cassava brown streak virus (UCBSV) and two predominant DNA viruses; African cassava mosaic virus (ACMV) and East African cassava mosaic virus (EACMV), which cause the economically important cassava brown streak disease (CBSD) and cassava mosaic disease (CMD) respectively. Our method, developed by analysing PCR products of viruses, was highly sensitive to detect target viruses from very low quantities of 4–10 femtograms. Multiplexing did not diminish sensitivity or accuracy compared to uniplex alternatives. The assay reliably detected and quantified four cassava viruses in field samples where CBSV and UCBSV synergy was observed in majority of mixed-infected varieties. Conclusions: We have developed a high-throughput qPCR diagnostic assay capable of specific and sensitive quantification of predominant DNA and RNA viruses of cassava in eastern Africa. Significance and Impact of the Study: The qPCR methods are a great improvement on the existing methods and can be used for monitoring virus spread as well as for accurate evaluation of the cassava varieties for virus resistance.
dc.format.extent1346–1356
dc.language.isoen
dc.subjectCassava
dc.subjectDna
dc.subjectPlant Diseases
dc.subjectRna
dc.subjectPcr
dc.titleHigh-throughput multiplex real-time PCR assay for the simultaneous quantification of DNA and RNA viruses infecting cassava plants
dc.typeJournal Article
dc.description.versionPeer Review
cg.contributor.crpRoots, Tubers and Bananas
cg.contributor.affiliationUniversity of Greenwich
cg.contributor.affiliationNaliendele Agricultural Research Institute, Tanzania
cg.contributor.affiliationInternational Institute of Tropical Agriculture
cg.coverage.regionAfrica
cg.coverage.regionEast Africa
cg.coverage.regionSouthern Africa
cg.coverage.countryBurundi
cg.coverage.countryCongo, Dr
cg.coverage.countryKenya
cg.coverage.countryMalawi
cg.coverage.countryMozambique
cg.coverage.countryUganda
cg.isijournalISI Journal
cg.authorship.typesCGIAR and developing country institute
cg.iitasubjectCassava
cg.iitasubjectPlant Diseases
cg.journalJournal of Applied Microbiology
cg.howpublishedFormally Published
cg.accessibilitystatusLimited Access
local.dspaceid78512
cg.targetaudienceScientists
cg.identifier.doihttps://dx.doi.org/10.1111/jam.13043


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