Cultivation of PCV2 in swine testicle cells using the shell vial technique and monitoring of viral replication by qPCR and RT-qPCR

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dc.contributor.authorCruz, Tais F. [UNESP]
dc.contributor.authorAraujo, Joao P. [UNESP]
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2014-12-03T13:10:54Z
dc.date.available2014-12-03T13:10:54Z
dc.date.issued2014-02-01
dc.description.abstractPorcine circovirus type 2 (PCV2) is difficult to isolate. Currently, no published articles have used the shell vial technique to isolate PCV2. In addition, the action of D-glucosamine on swine testicle cells (ST) has not been evaluated properly. Thus, the aim of this study was to determine an optimal concentration of D-glucosamine and to test the shell vial technique for PCV2 propagation in ST cells. The optimal concentration of D-glucosamine was determined to be 100 mM. Because PCV2 is noncytopathic, the traditional adsorption was compared to the shell vial technique for 15 passages by qPCR, and RT-qPCR for passages 12 through 15. The quantities of viral DNA (P=0.013) and ORF1-mRNA detected with the shell vial technique were two-fold higher than the obtained with traditional adsorption. The levels of ORF2-mRNA were similar for both methods; however, by passage 15, a six-fold increase in levels was observed with the shell vial technique. Therefore, the shell vial technique was more efficient for the cultivation of PCV2, and qPCR/RT-qPCR can be used to monitor viral replication. In addition, a high viral load (>2.7 x 10(10) DNA copies/ml) and high levels of viral mRNA expression indicated that the ST cells were persistently infected. (C) 2013 Elsevier B.V. All rights reserved.en
dc.description.affiliationUniv Estadual Paulista UNESP, Inst Biociencias, Dept Microbiol & Immunol, BR-18618970 Botucatu, SP, Brazil
dc.description.affiliationUnespUniv Estadual Paulista UNESP, Inst Biociencias, Dept Microbiol & Immunol, BR-18618970 Botucatu, SP, Brazil
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipIdFAPESP: 06/57976-6
dc.description.sponsorshipIdFAPESP: 06/59002-9
dc.description.sponsorshipIdCNPq: 471070/2007-6
dc.description.sponsorshipIdCNPq: 500905/2007-0
dc.description.sponsorshipIdCNPq: 12748/2008-6
dc.format.extent82-85
dc.identifierhttp://dx.doi.org/10.1016/j.jviromet.2013.10.025
dc.identifier.citationJournal Of Virological Methods. Amsterdam: Elsevier Science Bv, v. 196, p. 82-85, 2014.
dc.identifier.doi10.1016/j.jviromet.2013.10.025
dc.identifier.issn0166-0934
dc.identifier.urihttp://hdl.handle.net/11449/112635
dc.identifier.wosWOS:000336340300014
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofJournal of Virological Methods
dc.relation.ispartofjcr1.756
dc.relation.ispartofsjr0,858
dc.rights.accessRightsAcesso restrito
dc.sourceWeb of Science
dc.subjectPorcine circovirus 2en
dc.subjectSwine testicle cellsen
dc.subjectQuantitative PCRen
dc.subjectShell vial techniqueen
dc.subjectCentrifugal enhancementen
dc.titleCultivation of PCV2 in swine testicle cells using the shell vial technique and monitoring of viral replication by qPCR and RT-qPCRen
dc.typeArtigo
dcterms.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dcterms.rightsHolderElsevier B.V.
unesp.author.orcid0000-0002-9153-1485[2]
unesp.campusUniversidade Estadual Paulista (Unesp), Instituto de Biociências, Botucatupt

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