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Analytical validation of two RT-qPCR tests and detection of spring viremia of carp virus (SVCV) in persistently infected koi Cyprinus carpio

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dc.contributor.authorClouthier, Sharon C.en
dc.contributor.authorSchroeder, Tamaraen
dc.contributor.authorBueren, Emma K.en
dc.contributor.authorAnderson, Eric D.en
dc.contributor.authorEmmenegger, Evelineen
dc.contributor.departmentBiological Sciencesen
dc.date.accessioned2021-06-08T15:40:57Zen
dc.date.available2021-06-08T15:40:57Zen
dc.date.issued2021-02-25en
dc.description.abstractSpring viremia of carp virus (SVCV) ia a carp sprivivirus and a member of the genus Sprivivirus within the family Rhabdoviridae. The virus is the etiological agent of spring viremia of carp, a disease of cyprinid species including koi Cyprinus carpio L. and notifiable to the World Organisation for Animal Health. The goal of this study was to explore hypotheses regarding intergenogroup (Ia to Id) SVCV infection dynamics in juvenile koi and contemporaneously create new reverse-transcription quantitative PCR (RT-qPCR) assays and validate their analytical sensitivity, specificity (ASp) and repeatability for diagnostic detection of SVCV. RT-qPCR diagnostic tests targeting the SVCV nucleoprotein (Q2N) or glycoprotein (Q1G) nucleotides were pan-specific for isolates typed to SVCV genogroups Ia to Id. The Q2N test had broader ASp than Q1G because Q1G did not detect SVCV isolate 20120450 and Q2N displayed occasional detection of pike fry sprivivirus isolate V76. Neither test cross-reacted with other rhabdoviruses, infectious pancreatic necrosis virus or co-localizing cyprinid herpesvirus 3. Both tests were sensitive with observed 50% limits of detection of 3 plasmid copies and high repeatability. Test analysis of koi immersed in SVCV showed that the virus could be detected for at least 167 d following exposure and that titer, prevalence, replicative rate and persistence in koi were correlated significantly with virus virulence. In this context, high virulence SVCV isolates were more prevalent, reached higher titers quicker and persisted in koi for longer periods of time relative to moderate and low virulence isolates.en
dc.description.notesThis study was supported by the Centre of Expertise for Aquatic Animal Health Research & Development (Fisheries and Oceans Canada) and the Emerging Diseases Cyclical Fund (US Geological Survey). We thank Ron Hedrick and Tomo Kurobe for CyHV-3 isolate F98-50; Kyle Garver for the VHSV and IHNV isolates; and David Stone, Andrew Goodwin, Peng Jia and Hong Liu for the carp and pike fry sprivivirus isolates. Any use of trade, firm or product names is for descriptive purposes only and does not imply endorsement by the US government.en
dc.description.sponsorshipCentre of Expertise for Aquatic Animal Health Research & Development (Fisheries and Oceans Canada); Emerging Diseases Cyclical Fund (US Geological Survey)en
dc.description.versionPublished versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.3354/dao03564en
dc.identifier.eissn1616-1580en
dc.identifier.issn0177-5103en
dc.identifier.pmid33629660en
dc.identifier.urihttp://hdl.handle.net/10919/103699en
dc.identifier.volume143en
dc.language.isoenen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectSVCVen
dc.subjectRT-qPCRen
dc.subjectAnalytical specificityen
dc.subjectAnalytical sensitivityen
dc.subjectPersistent infectionen
dc.subjectKoien
dc.subjectSprivivirusen
dc.titleAnalytical validation of two RT-qPCR tests and detection of spring viremia of carp virus (SVCV) in persistently infected koi Cyprinus carpioen
dc.title.serialDiseases of Aquatic Organismsen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
dc.type.dcmitypeStillImageen

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