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Asymmetric PCR ELISA: increased sensitivity and reduced costs for the detection of plant viruses

2002Journal article Restricted
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dc.contributor.authorNolasco, Gustavo
dc.contributor.authorSequeira, Z.
dc.contributor.authorSoares, C.
dc.contributor.authorMansinho, A.
dc.contributor.authorBailey, A. M.
dc.contributor.authorNiblett, C. l.
dc.date.accessioned2015-06-15T14:13:47Z
dc.date.available2015-06-15T14:13:47Z
dc.date.issued2002
dc.description.abstractPCR ELISA is the immunodetection of the products of a polymerase chain reaction (PCR). It is effective for detecting and differentiating plant viral nucleic acids, but as currently performed, it is laborious and expensive. The procedure has been modified and simplified by using asymmetric PCR. This eliminated the need to denature and neutralize samples prior to hybridization. It also increased the relative concentration of the target DNA species, making PCR ELISA more sensitive than TaqMan(TM), a fluorescence-based detection method. Reducing the reaction volumes to half and the concentration of the dNTPs and the digoxigenin label by tenfold significantly reduced the costs of PCR ELISA without reducing its sensitivity. The usefulness of these modifications was demonstrated for the detection of Citrus tristeza virus and Rupestris stem pitting-associated virus. We expect that with only minor modifications asymmetric PCR ELISA could be used effectively for the detection of most nucleic acid molecules of interest.
dc.identifier.doihttps://dx.doi.org/10.1023/A:1015649429160
dc.identifier.issn0929-1873
dc.identifier.urihttp://hdl.handle.net/10400.1/6243
dc.language.isoeng
dc.peerreviewedyes
dc.publisherKluwer Academic Publ.
dc.relation.isbasedonP-000-PF7
dc.titleAsymmetric PCR ELISA: increased sensitivity and reduced costs for the detection of plant viruses
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage298
oaire.citation.startPage293
oaire.citation.titleEuropean Journal of Plant Pathology
oaire.citation.volume108
person.familyNameNolasco
person.familyNameSOARES
person.givenNameGustavo
person.givenNameCRISTIANO
person.identifier.orcid0000-0002-7025-6023
person.identifier.orcid0000-0002-2789-8506
person.identifier.scopus-author-id6602625528
rcaap.rightsrestrictedAccess
rcaap.typearticle
relation.isAuthorOfPublication87a8035f-bc15-44dd-b8c3-4288c5660fff
relation.isAuthorOfPublication21359474-b0e6-4d62-a0b1-fa87a750b1ca
relation.isAuthorOfPublication.latestForDiscovery87a8035f-bc15-44dd-b8c3-4288c5660fff

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