Development of a PCR-based assay for detection, quantification, and genotyping of human adenoviruses.
作者： Barbara Chmielewicz , Andreas Nitsche , Brunhilde Schweiger , Heinz Ellerbrok
DOI: 10.1373/CLINCHEM.2004.045088
关键词: Polymerase chain reaction 、 Genotype 、 Virus quantification 、 Nested polymerase chain reaction 、 Biology 、 Genotyping 、 Transplantation 、 DNA polymerase 、 Genome 、 Molecular biology
摘要: Background: Adenoviruses (AdVs) can cause serious disease in immunosuppressed patients, particularly those undergoing allogeneic stem cell transplantation. A method for virus quantification clinical specimens is essential monitoring patient adenoviral loads and evaluating new therapeutic approaches. Methods: We developed a PCR-based assay that combines detection genotyping of human AdVs, targeting highly conserved region the genome coding DNA polymerase (AdV DPol PCR). tested diagnostic applicability this by analyzing 159 from children with respiratory comparing results obtained nested PCR analysis. Results: The detected all currently known AdV serotypes, limit ∼10 equivalents per reaction 49 51 serotypes. No cross-reactivity to or other viruses was observed. In addition, PCR-positive samples achieved within minutes fluorescence curve melting analysis LightCycler instrument using 6 pairs hybridization probes, each specific single species. Results were good concordance PCR. Conclusion: presented suitable tool AdVs samples.
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