A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers

dc.contributorUniversidade Estadual Paulista (UNESP)
dc.creatorCiapina, L. P.
dc.creatorAlves, LMC
dc.creatorLemos, EGM
dc.date2014-05-20T13:17:29Z
dc.date2014-05-20T13:17:29Z
dc.date2004-01-01
dc.date.accessioned2017-04-05T19:38:13Z
dc.date.available2017-04-05T19:38:13Z
dc.identifierJournal of Applied Microbiology. Oxford: Blackwell Publishing Ltd, v. 96, n. 3, p. 546-551, 2004.
dc.identifier1364-5072
dc.identifierhttp://hdl.handle.net/11449/3927
dc.identifier10.1111/j.1365-2672.2004.02176.x
dc.identifierWOS:000188990400014
dc.identifierWOS000188990400014.pdf
dc.identifierhttp://dx.doi.org/10.1111/j.1365-2672.2004.02176.x
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/853406
dc.descriptionAims: Detection of Xylella fastidiosa in citrus plants and insect vectors.Methods and Results: Chelex 100 resin matrix was successfully standardized allowing a fast DNA extraction of X. fastidiosa. An amplicon of 500 bp was observed in samples of citrus leaf and citrus xylem extract, with and without symptoms of citrus variegated chlorosis, using PCR with a specific primer set indicating the presence of X. fastidiosa. The addition of insoluble acid-washed polyvinylpyrrolidone (PVPP) prior to DNA extraction of insect samples using Chelex 100 resin together with nested-PCR permitted the detection of X. fastidiosa within sharpshooter heads with great sensitivity. It was possible to detect up to two bacteria per reaction. From 250 sharpshooter samples comprising four species (Dilobopterus costalimai, Oncometopia facialis, Bucephalogonia xanthopis and Acrogonia sp.), 87 individuals showed positive results for X. fastidiosa in a nested-PCR assay.Conclusions: the use of Chelex 100 resin allowed a fast and efficient DNA extraction to be used in the detection of X. fastidiosa in citrus plants and insect vectors by PCR and nested-PCR assays, respectively.Significance and Impact of the study: the employment of efficient and sensitive methods to detect X. fastidiosa in citrus plants and insect vectors will greatly assist epidemiological studies.
dc.languageeng
dc.publisherBlackwell Publishing
dc.relationJournal of Applied Microbiology
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectChelex 100
dc.subjectcitrus variegated chlorosis
dc.subjectDNA extraction
dc.subjectNested PCR
dc.subjectsharpshooter
dc.subjectXylella fastidiosa
dc.titleA nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers
dc.typeOtro


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