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Development of duplex TaqMan-based real-time PCR assay for the simultaneous detection of Perkinsus olseni and P. chesapeaki in host Manila clam tissue samples

dc.rights.licenseopenen_US
dc.contributor.authorITOIZ, Sarah
dc.contributor.authorPERENNOU, Morgan
dc.contributor.authorMOURONVALLE, Clara
dc.contributor.authorDERELLE, Evelyne
dc.contributor.authorLE GOIC, Nelly
dc.contributor.authorBIDAULT, Adeline
hal.structure.identifierEnvironnements et Paléoenvironnements OCéaniques [EPOC]
dc.contributor.authorDE MONTAUDOUIN, Xavier
IDREF: 075931664
dc.contributor.authorARZUL, Isabelle
dc.contributor.authorSOUDANT, Philippe
dc.contributor.authorCHAMBOUVET, Aurelie
dc.date.accessioned2023-09-11T09:42:37Z
dc.date.available2023-09-11T09:42:37Z
dc.date.issued2021-09-01
dc.identifier.issn0022-2011en_US
dc.identifier.urioai:crossref.org:10.1016/j.jip.2021.107603
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/183645
dc.description.abstractEnThe aetiological agent Perkinsus olseni is globally recognised as a major threat for shellfish production considering its wide geographical distribution across Asia, Europe, Australia and South America. Another species, Perkinsus chesapeaki, which has never been known to be associated with significant mortality events, was recently detected along French coasts infecting clam populations sporadically in association with P. olseni. Identifying potential cryptic infections affecting Ruditapes philippinarum is essential to develop appropriate host resource management strategies. Here, we developed a molecular method based on duplex real-time quantitative PCR for the simultaneous detection of these two parasites, P. olseni and P. chesapeaki, in the different clam tissues: gills, digestive gland, foot, mantle, adductor muscle and the rest of the soft body. We firstly checked the presence of possible PCR inhibitors in host tissue samples. The qPCR reactions were inhibited depending on the nature of the host organ. The mantle and the rest of the soft body have a high inhibitory effect from threshold of host gDNA concentration of 2 ng.µL−1, the adductor muscle and the foot have an intermediate inhibition of 5 ng.µL−1, and the gills and digestive gland do not show any inhibition of the qPCR reaction even at the highest host gDNA concentration of 20 ng.µL−1. Then, using the gills as a template, the suitability of the molecular technique was checked in comparison with the Ray’s Fluid Thioglycolate Medium methodology recommended by the World Organisation for Animal Health. The duplex qPCR method brought new insights and unveiled cryptic infections as the co-occurrence of P. olseni and P. chesapeaki from in situ tissue samples in contrast to the RFTM diagnosis. The development of this duplex qPCR method is a fundamental work to monitor in situ co-infections that will lead to optimised resource management and conservation strategies to deal with emerging diseases.
dc.language.isoENen_US
dc.rightsAttribution-NonCommercial 3.0 United States*
dc.rights.urihttp://creativecommons.org/licenses/by-nc/3.0/us/*
dc.sourcecrossref
dc.title.enDevelopment of duplex TaqMan-based real-time PCR assay for the simultaneous detection of Perkinsus olseni and P. chesapeaki in host Manila clam tissue samples
dc.typeArticle de revueen_US
dc.identifier.doi10.1016/j.jip.2021.107603en_US
dc.subject.halSciences de l'environnementen_US
bordeaux.journalJournal of Invertebrate Pathologyen_US
bordeaux.page107603en_US
bordeaux.volume184en_US
bordeaux.hal.laboratoriesEPOC : Environnements et Paléoenvironnements Océaniques et Continentaux - UMR 5805en_US
bordeaux.institutionUniversité de Bordeauxen_US
bordeaux.institutionCNRSen_US
bordeaux.teamECOBIOCen_US
bordeaux.peerReviewedouien_US
bordeaux.inpressnonen_US
bordeaux.import.sourcedissemin
hal.popularnonen_US
hal.audienceInternationaleen_US
hal.exportfalse
workflow.import.sourcedissemin
dc.rights.ccPas de Licence CCen_US
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