dc.contributor.advisor | Weidmann, Manfred PD Dr. | de |
dc.contributor.author | Ehnts, Kai Ilmo | de |
dc.date.accessioned | 2013-07-19T08:51:46Z | de |
dc.date.available | 2013-08-13T22:50:04Z | de |
dc.date.issued | 2013-07-19 | de |
dc.identifier.uri | http://hdl.handle.net/11858/00-1735-0000-0001-BAD4-F | de |
dc.identifier.uri | http://dx.doi.org/10.53846/goediss-3943 | |
dc.language.iso | deu | de |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/3.0/ | |
dc.subject.ddc | 610 | de |
dc.title | Entwicklung von Rekombinase-Polymerase-Amplifikations-Nachweisverfahren für virale Erreger von Atemwegsinfektionen | de |
dc.type | doctoralThesis | de |
dc.title.translated | Development of a panel of recombinase polymerase amplification assays for detection of respiratory viruses | de |
dc.contributor.referee | Hufert, Frank Torsten Prof. Dr. | de |
dc.date.examination | 2013-08-06 | de |
dc.description.abstracteng | Objective:
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.coReferee | Bartels, Iris Prof. Dr. | de |
dc.contributor.thirdReferee | Oppermann, Martin Prof. Dr. | de |
dc.subject.ger | RPA | de |
dc.subject.ger | Rekombinase-Polymerase-Amplifikation | de |
dc.subject.ger | Influenza | de |
dc.subject.ger | Adenovirus | de |
dc.subject.ger | Schnelltest | de |
dc.subject.ger | Nukleinsäurenachweisverfahren | de |
dc.subject.ger | Grippe | de |
dc.subject.ger | Atemwegsinfektion | de |
dc.subject.eng | RPA | de |
dc.subject.eng | recombinase polymerase amplification | de |
dc.subject.eng | Influenza | de |
dc.subject.eng | Adenovirus | de |
dc.subject.eng | PoC | de |
dc.subject.eng | point-of-care diagnostic | de |
dc.subject.eng | detection of nucleic acid | de |
dc.subject.eng | PCR | de |
dc.subject.eng | Nucleic Acid Amplification Test | de |
dc.subject.eng | NAAT | de |
dc.subject.eng | Flu | de |
dc.subject.eng | Acute respiratory infection | de |
dc.identifier.urn | urn:nbn:de:gbv:7-11858/00-1735-0000-0001-BAD4-F-9 | de |
dc.affiliation.institute | Medizinische Fakultät | de |
dc.subject.gokfull | Medizin (PPN619874732) | de |
dc.subject.gokfull | Labormedizin / Klinische Chemie - Allgemein- und Gesamtdarstellungen (PPN61987564X) | de |
dc.subject.gokfull | Diagnostik {Medizin} (PPN619875739) | de |
dc.subject.gokfull | Medizinische Mikrobiologie / Medizinische Virologie / Medizinische Mykologie / Infektionskrankheiten / Hygiene / Impfung / Parasitologie / Tropenmedizin - Allgemein- und Gesamtdarstellungen (PPN619875356) | de |
dc.description.embargoed | 2013-08-13 | de |
dc.identifier.ppn | 755421957 | de |