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Development of a PCR-ELISA assay for diagnosis of Perkinsus marinus and Perkinsus atlanticus infections in bivalve molluscs

2004Journal article Restricted access
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dc.contributor.authorElandalloussi, L. M.
dc.contributor.authorLeite, Ricardo
dc.contributor.authorAfonso, R.
dc.contributor.authorNunes, Patrícia A.
dc.contributor.authorRobledo, J. A. F.
dc.contributor.authorVasta, G. R.
dc.contributor.authorCancela, Leonor
dc.date.accessioned2015-11-30T15:17:51Z
dc.date.available2015-11-30T15:17:51Z
dc.date.issued2004
dc.description.abstractPerkinsus atlanticus and P. marinus have been associated with mass mortality of bivalve molluscs. Perkinsus infections are routinely diagnosed by histology or the fluid thioglycollate medium (FTM) assay. In this study, we describe the development of a PCR-enzyme-linked immunosorbent assay (ELISA) for amplification and rapid detection of Perkinsus species. The PCR reactions were selected to either amplify an IGS sequence region shared by currently accepted Perkinsus species or to simultaneously amplify IGS regions specific to either P. atlanticus or P. marinus. The specific hybridisation of DIG-labelled amplified products to species-specific capture probes was detected colorimetrically. This assay is able to specifically detect P. atlanticus and P. marinus, and the intensity of the colorimetric signal is dependent upon the amount of amplified product. The PCR-ELISA assay format is 100-fold more sensitive than visualisation of PCR products on ethidium bromide (EtdBr)-stained agarose gels, and as sensitive as Southern hybridisation. The sensitivity limit of PCR-ELISA was 1 pg of DNA front P. atlanticus. No cross-reactivity of the assay was observed against the host DNA. When applied to the detection of P. atlanticus in clams, 39 samples out of 45 yielded concordant results for FTM assay and PCR-ELISA detection. (C) 2003 Elsevier Ltd. All rights reserved.
dc.identifier.doihttps://dx.doi.org/10.1016/j.mcp.2003.09.008
dc.identifier.issn0890-8508
dc.identifier.otherAUT: PCA01721; LCA00739;
dc.identifier.urihttp://hdl.handle.net/10400.1/7219
dc.language.isoeng
dc.peerreviewedyes
dc.publisherElsevier
dc.relation.isbasedonP-000-B2C
dc.titleDevelopment of a PCR-ELISA assay for diagnosis of Perkinsus marinus and Perkinsus atlanticus infections in bivalve molluscs
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage96
oaire.citation.startPage89
oaire.citation.titleMolecular and Cellular Probes
oaire.citation.volume18
person.familyNameLeite
person.familyNameAfonso
person.familyNameNunes Cabrita
person.familyNameCancela
person.givenNameRicardo
person.givenNameRicardo
person.givenNamePatrícia
person.givenNameM. Leonor
person.identifier447622
person.identifier.ciencia-idF115-256E-C84D
person.identifier.orcid0000-0002-9622-3895
person.identifier.orcid0000-0002-3884-9312
person.identifier.orcid0000-0002-7148-8950
person.identifier.orcid0000-0003-3114-6662
person.identifier.ridB-3389-2008
person.identifier.scopus-author-id8362650900
person.identifier.scopus-author-id7006823084
rcaap.rightsrestrictedAccess
rcaap.typearticle
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relation.isAuthorOfPublication30644154-3327-45af-a3ed-f5e221951b4c
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