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Entwicklung von Rekombinase-Polymerase-Amplifikations-Nachweisverfahren für virale Erreger von Atemwegsinfektionen

dc.contributor.advisorWeidmann, Manfred PD Dr.de
dc.contributor.authorEhnts, Kai Ilmode
dc.date.accessioned2013-07-19T08:51:46Zde
dc.date.available2013-08-13T22:50:04Zde
dc.date.issued2013-07-19de
dc.identifier.urihttp://hdl.handle.net/11858/00-1735-0000-0001-BAD4-Fde
dc.identifier.urihttp://dx.doi.org/10.53846/goediss-3943
dc.language.isodeude
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/
dc.subject.ddc610de
dc.titleEntwicklung von Rekombinase-Polymerase-Amplifikations-Nachweisverfahren für virale Erreger von Atemwegsinfektionende
dc.typedoctoralThesisde
dc.title.translatedDevelopment of a panel of recombinase polymerase amplification assays for detection of respiratory virusesde
dc.contributor.refereeHufert, Frank Torsten Prof. Dr.de
dc.date.examination2013-08-06de
dc.description.abstractengObjective: Acute respiratory infections (ARI), ranging from the common cold to life threatening Influenza, are frequent causes for medical consultations. Point-of-Care (PoC) diagnosis of Influenza facilitates early antiviral treatment and can lead to a reduction in antibiotic use. We designed three assays for the fast and sensitive isothermal PoC detection of Influenza A, Influenza B and Adenovirus using Recombinase-Polymerase-Amplification. Methods: Following existing PCR amplicons, we aligned several random Genbank sequences and designed specific primer and probe sets for Influenza A, Influenza B and Adenoviruses. We developed quantitative standards of synthetic viral RNA and DNA respectively, which where calibrated using real-time PCR. For human Adenoviruses three standard preparations of serotype 1, 4 and 7 were developed to cover those serotypes causing severe respiratory tract infections. Sensitivity of each assay was assessed in 8 similar runs in a ESEQuant Tube Scanner instrument for 20 minutes at 42°C. The obtained data was statistically analysed via Probit analysis. Results: The number of molecules detected in 95% of test runs in a 50 µl volume were as per Probit analysis: Influenza-A 218 copies, Influenza-B 131 copies, HAdV-1 12 copies, HAdV-4 131 copies, HAdV-7 16 copies. The runtime until positive results was for Influenza-A not exceeding 9 minutes, for Influenza-B not exceeding 8 minutes and for Adenoviruses not exceeding 14 minutes. Conclusion: Recombinase-Polymerase-Amplification provides a very fast, sensitive and specific way of isothermal detection of respiratory viruses. These assays can be easily adapted to microfluidic Point-of-Care devices.de
dc.contributor.coRefereeBartels, Iris Prof. Dr.de
dc.contributor.thirdRefereeOppermann, Martin Prof. Dr.de
dc.subject.gerRPAde
dc.subject.gerRekombinase-Polymerase-Amplifikationde
dc.subject.gerInfluenzade
dc.subject.gerAdenovirusde
dc.subject.gerSchnelltestde
dc.subject.gerNukleinsäurenachweisverfahrende
dc.subject.gerGrippede
dc.subject.gerAtemwegsinfektionde
dc.subject.engRPAde
dc.subject.engrecombinase polymerase amplificationde
dc.subject.engInfluenzade
dc.subject.engAdenovirusde
dc.subject.engPoCde
dc.subject.engpoint-of-care diagnosticde
dc.subject.engdetection of nucleic acidde
dc.subject.engPCRde
dc.subject.engNucleic Acid Amplification Testde
dc.subject.engNAATde
dc.subject.engFlude
dc.subject.engAcute respiratory infectionde
dc.identifier.urnurn:nbn:de:gbv:7-11858/00-1735-0000-0001-BAD4-F-9de
dc.affiliation.instituteMedizinische Fakultätde
dc.subject.gokfullMedizin (PPN619874732)de
dc.subject.gokfullLabormedizin / Klinische Chemie - Allgemein- und Gesamtdarstellungen (PPN61987564X)de
dc.subject.gokfullDiagnostik {Medizin} (PPN619875739)de
dc.subject.gokfullMedizinische Mikrobiologie / Medizinische Virologie / Medizinische Mykologie / Infektionskrankheiten / Hygiene / Impfung / Parasitologie / Tropenmedizin - Allgemein- und Gesamtdarstellungen (PPN619875356)de
dc.description.embargoed2013-08-13de
dc.identifier.ppn755421957de


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