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2011-02-10Zeitschriftenartikel DOI: 10.1186/1743-422X-8-63
Highly sensitive detection of the group A Rotavirus using Apolipoprotein H-coated ELISA plates compared to quantitative real-time PCR
dc.contributor.authorAdlhoch, Cornelia
dc.contributor.authorKaiser, Marco
dc.contributor.authorHoehne, Marina
dc.contributor.authorMarques, Andreas Mas
dc.contributor.authorStefas, Ilias
dc.contributor.authorVeas, Francisco
dc.contributor.authorEllerbrok, Heinz
dc.date.accessioned2018-05-07T14:26:05Z
dc.date.available2018-05-07T14:26:05Z
dc.date.created2011-02-23
dc.date.issued2011-02-10none
dc.identifier.otherhttp://edoc.rki.de/oa/articles/reCo9XafkP0pw/PDF/27XyMtpvwZLk.pdf
dc.identifier.urihttp://edoc.rki.de/176904/819
dc.description.abstractBackground: The principle of a capture ELISA is binding of specific capture antibodies (polyclonal or monoclonal) to the surface of a suitable 96 well plate. These immobilized antibodies are capable of specifically binding a virus present in a clinical sample. Subsequently, the captured virus is detected using a specific detection antibody. The drawback of this method is that a capture ELISA can only function for a single virus captured by the primary antibody. Human Apolipoprotein H (ApoH) or b2-glycoprotein 1 is able to poly-specifically bind viral pathogens. Replacing specific capture antibodies by ApoH should allow poly-specific capture of different viruses that subsequently could be revealed using specific detection antibodies. Thus, using a single capture ELISA format different viruses could be analysed depending on the detection antibody that is applied. In order to demonstrate that this is a valid approach we show detection of group A rotaviruses from stool samples as a proof of principle for a new method of capture ELISA that should also be applicable to other viruses. Results: Stool samples of different circulating common human and potentially zoonotic group A rotavirus strains, which were pretested in commercial EIAs and genotyped by PCR, were tested in parallel in an ApoH-ELISA set-up and by quantitative real-time PCR (qPCR). Several control samples were included in the analysis. The ApoH-ELISA was suitable for the capture of rotavirus-particles and the detection down to 1,000 infectious units (TCID50/ml). Subsets of diagnostic samples of different G- and P-types were tested positive in the ApoH-ELISA in different dilutions. Compared to the qPCR results, the analysis showed high sensitivity, specificity and low cross-reactivity for the ApoH-ELISA, which was confirmed in receiver operating characteristics (ROC) analysis. Conclusions: In this study the development of a highly sensitive and specific capture ELISA was demonstrated by combining a poly-specific ApoH capture step with specific detection antibodies using group A rotaviruses as an example.eng
dc.language.isoeng
dc.publisherRobert Koch-Institut, Infektionsepidemiologie; Robert Koch-Institut, Biologische Sicherheit
dc.subjectAdulteng
dc.subjectHumanseng
dc.subjectSensitivity and Specificityeng
dc.subjectInfanteng
dc.subjectPolymerase Chain Reaction/methodseng
dc.subjectEnzyme-Linked Immunosorbent Assay/methodseng
dc.subjectFeces/virologyeng
dc.subjectVirology/methodseng
dc.subjectRotavirus/isolation & purificationeng
dc.subjectRotavirus Infections/diagnosiseng
dc.subjectSerologic Tests/methodseng
dc.subjectbeta 2-Glycoprotein I/diagnostic useeng
dc.subject.ddc610 Medizin
dc.titleHighly sensitive detection of the group A Rotavirus using Apolipoprotein H-coated ELISA plates compared to quantitative real-time PCR
dc.typeperiodicalPart
dc.identifier.urnurn:nbn:de:0257-10012627
dc.identifier.doi10.1186/1743-422X-8-63
dc.identifier.doihttp://dx.doi.org/10.25646/744
local.edoc.container-titleVirology Journal
local.edoc.fp-subtypeArtikel
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttp://www.virologyj.com/content/8/1/63
local.edoc.container-publisher-nameBioMedCentral
local.edoc.container-volume8
local.edoc.container-issue63
local.edoc.container-year2011

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