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2021-06-02Zeitschriftenartikel DOI: 10.25646/8659
Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2
dc.contributor.authorMichel, Janine
dc.contributor.authorNeumann, Markus
dc.contributor.authorKrause, Eva
dc.contributor.authorRinner, Thomas
dc.contributor.authorMuzeniek, Therese
dc.contributor.authorGrossegesse, Marica
dc.contributor.authorHille, Georg
dc.contributor.authorSchwarz, Franziska
dc.contributor.authorPuyskens, Andreas
dc.contributor.authorFörster, Sophie
dc.contributor.authorBiere, Barbara
dc.contributor.authorBourquain, Daniel
dc.contributor.authorDomingo, Cristina
dc.contributor.authorBrinkmann, Annika
dc.contributor.authorSchaade, Lars
dc.contributor.authorSchrick, Livia
dc.contributor.authorNitsche, Andreas
dc.date.accessioned2021-06-16T07:27:23Z
dc.date.available2021-06-16T07:27:23Z
dc.date.issued2021-06-02none
dc.identifier.other10.1186/s12985-021-01559-3
dc.identifier.urihttp://edoc.rki.de/176904/8379
dc.description.abstractBackground The reliable detection of SARS-CoV-2 has become one of the most important contributions to COVID-19 crisis management. With the publication of the first sequences of SARS-CoV-2, several diagnostic PCR assays have been developed and published. In addition to in-house assays the market was flooded with numerous commercially available ready-to-use PCR kits, with both approaches showing alarming shortages in reagent supply. Aim Here we present a resource-efficient in-house protocol for the PCR detection of SARS- CoV-2 RNA in patient specimens (RKI/ZBS1 SARS-CoV-2 protocol). Methods Two duplex one-step real-time RT-PCR assays are run simultaneously and provide information on two different SARS-CoV-2 genomic regions. Each one is duplexed with a control that either indicates potential PCR inhibition or proves the successful extraction of nucleic acid from the clinical specimen. Results Limit of RNA detection for both SARS-CoV-2 assays is below 10 genomes per reaction. The protocol enables testing specimens in duplicate across the two different SARS-CoV-2 PCR assays, saving reagents by increasing testing capacity. The protocol can be run on various PCR cyclers with several PCR master mix kits. Conclusion The presented RKI/ZBS1 SARS-CoV-2 protocol represents a cost-effective alternative in times of shortages when commercially available ready-to-use kits may not be available or affordable.eng
dc.language.isoengnone
dc.publisherRobert Koch-Institut
dc.rights(CC BY 3.0 DE) Namensnennung 3.0 Deutschlandger
dc.rights.urihttp://creativecommons.org/licenses/by/3.0/de/
dc.subjectSARS-CoV-2eng
dc.subjectCOVID-19eng
dc.subjectDiagnosticseng
dc.subjectReal-time PCReng
dc.subjectInternal controleng
dc.subject.ddc610 Medizin und Gesundheitnone
dc.titleResource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2none
dc.typearticle
dc.identifier.urnurn:nbn:de:kobv:0257-176904/8379-1
dc.identifier.doihttp://dx.doi.org/10.25646/8659
dc.type.versionpublishedVersionnone
local.edoc.container-titleVirology Journalnone
local.edoc.container-issn1743-422Xnone
local.edoc.pages11none
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttps://virologyj.biomedcentral.com/articles/10.1186/s12985-021-01559-3none
local.edoc.container-publisher-nameSpringer Naturenone
local.edoc.container-volume18none
local.edoc.rki-departmentZentrum für Biologische Gefahren und Spezielle Pathogenenone
dc.description.versionPeer Reviewednone

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