Fabienne Rehren

Publications of Fabienne Rehren

• Unique sequence features of the Adenovirus 31 complete genomic sequence are conserved in clinical isolates.

  Authors: Soeren Hofmayer, Ijad Madisch, Sebastian Darr, Fabienne Rehren, Albert Heim

  BMC genomics. 11/2009; 10(1):557.

  ABSTRACT: BACKGROUND: Human adenoviruses (HAdV) are causing a broad spectrum of diseases. One of the most severe forms of adenovirus infection is a disseminated disease resulting in significantABSTRACT: BACKGROUND: Human adenoviruses (HAdV) are causing a broad spectrum of diseases. One of the most severe forms of adenovirus infection is a disseminated disease resulting in significant morbidity and mortality. Several reports in recent years have identified HAdV-31 from species A (HAdV-A31) as a cause of disseminated disease in children following haematopoetic stem cell transplantation (hSCT) and liver transplantation. We sequenced and analyzed the complete genome of the HAdV-A31 prototype strain to uncover unique sequence motifs associated with its high virulence. Moreover, we sequenced coding regions known to be essential for tropism and virulence (early transcription units E1A, E3, E4, the fiber knob and the penton base) of HAdV-A31 clinical isolates from patients with disseminated disease. RESULTS: The genome size of HAdV-A31 is 33763 base pairs (bp) in length with a GC content of 46.36%. Nucleotide alignment to the closely related HAdV-A12 revealed an overall homology of 84.2%. The genome organization into early, intermediate and late regions is similar to HAdV-A12. Sequence analysis of the prototype strain showed unique sequence features such as an immunoglobulin-like domain in the species A specific gene product E3 CR1 beta and a potentially integrin binding RGD motif in the C-terminal region of the protein IX. These features were conserved in all analyzed clinical isolates. Overall, amino acid sequences of clinical isolates were highly conserved compared to the prototype (99.2 to 100%), but a synonymous/non synonymous ratio (S/N) of 2.36 in E3 CR1-beta suggested positive selection. CONCLUSIONS: Unique sequence features of HAdV-A31 may enhance its ability to escape the host's immune surveillance and may facilitate a promiscuous tropism for various tissues. Moderate evolution of clinical isolates did not indicate the emergence of new HAdV-A31 subtypes in the recent years.

  Download

• Phylogeny and primary structure analysis of fiber shafts of all human adenovirus types for rational design of adenoviral gene therapy vectors.

  Authors: Sebastian Darr, Ijad Madisch, Sören Hofmayer, Fabienne Rehren, Albert Heim

  The Journal of general virology. 09/2009;

  The fiber shaft of human adenoviruses (HAdV) is essential for bringing the penton base in proximity to the secondary cellular receptor. Fiber shaft sequences of all 53 HAdV types were studied.The fiber shaft of human adenoviruses (HAdV) is essential for bringing the penton base in proximity to the secondary cellular receptor. Fiber shaft sequences of all 53 HAdV types were studied. Phylogeny of the fiber shaft revealed clustering corresponding to the HAdV species concept. An intraspecies recombination hot spot was found at the shaft/knob boundary, a highly conserved sequence stretch. For example, HAdV-D20 clustered with -D23 in the fiber shaft but with HAdV-D47 in the fiber knob. Although all shafts exhibited the typical pseudorepeats, sequence divergence was found to be as high as 92% interspecies and 54% intraspecies. In contrast to a previous study, a flexibility motif (KXGGLXFD/N) was found in eight HAdV-D types whereas the putative heparan sulfate binding site (KKTK) was only found in species HAdV-C. Our results suggest that pseudotyping of gene therapy vectors at the shaft/knob-boundary is feasible but flexibility data of shafts should be considered.

  Download

• Rapid routine detection of enterovirus RNA in cerebrospinal fluid by a one-step real-time RT-PCR assay.

  Authors: Uta Dierssen, Fabienne Rehren, Cornelia Henke-Gendo, Gabi Harste, Albert Heim

  Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology. 06/2008; 42(1):58-64.

  BACKGROUND AND OBJECTIVES: This study provides a one-step transcription/real-time (TaqMan probe) PCR assay (TM-PCR) with new consensus primer and probe sequences for generic detection of humanBACKGROUND AND OBJECTIVES: This study provides a one-step transcription/real-time (TaqMan probe) PCR assay (TM-PCR) with new consensus primer and probe sequences for generic detection of human pathogenic enteroviruses including difficult to detect ones like for instance Echovirus 30. The amplicon included parts of domain IV and V of the highly conserved internal ribosomal entry site. Generic detection was confirmed by testing a panel of 41 prototypes representing all five human enterovirus/poliovirus species. STUDY DESIGN AND RESULTS: The 95% detection limit was found to be 100 copies per run using in vitro transcribed coxsackievirus B3 RNA. TM-PCR was compared to an in house nested-PCR assay implemented in detecting enterovirus RNA from CSF samples of patients suffering from meningitis and encephalitis. Concordant results were obtained in all samples (11 positive, 101 negative). Specificity was confirmed with laboratory strains of other neurotropic viruses, and by testing 76 CSF samples of patients with encephalomyelitis disseminata, which all gave negative results. CONCLUSIONS: The new TM-PCR is a convincing alternative to conventional PCR protocols for the diagnosis of enterovirus meningitis. The one-step strategy limits hands on time and cross contamination risk combined with accelerated assay procedure of only 100 min.

• Unique sequence features of the Adenovirus 31 prototype complete genomic sequence are conserved in clinical isolates from cases of disseminated infection

  Authors: Sören Hofmayer, Ijad Madisch, Fabienne Rehren, Albert Heim

  01/2008

• Unique Sequence Features of the Human Adenovirus 31 Complete Genomic Sequence are Conserved in Clinical Isolates

  Authors: Soeren Hofmayer, Sebastian Darr, Fabienne Rehren, Albert Heim, Ijad Madisch

  Background: Human adenoviruses (HAdV) are causing a broad spectrum of diseases. One of the most severe forms of adenovirus infection is a disseminated disease resulting in significant morbidity andBackground: Human adenoviruses (HAdV) are causing a broad spectrum of diseases. One of the most severe forms of adenovirus infection is a disseminated disease resulting in significant morbidity and mortality. Several reports in recent years have identified HAdV-31 from species A (HAdV-A31) as a cause of disseminated disease in children following haematopoetic stem cell transplantation (hSCT) and liver transplantation. We sequenced and analyzed the complete genome of the HAdV-A31 prototype strain to uncover unique sequence motifs associated with its high virulence. Moreover, we sequenced coding regions known to be essential for tropism and virulence (early transcription units E1A, E3, E4, the fiber knob and the penton base) of HAdV-A31 clinical isolates from patients with disseminated disease. Results: The genome size of HAdV-A31 is 33763 base pairs (bp) in length with a GC content of 46.36%. Nucleotide alignment to the closely related HAdV-A12 revealed an overall homology of 84.2%. The genome organization into early, intermediate and late regions is similar to HAdV-A12. Sequence analysis of the prototype strain showed unique sequence features such as an immunoglobulin-like domain in the species A specific gene product E3 CR1 beta and a potentially integrin binding RGD motif in the C-terminal region of the protein IX. These features were conserved in all analyzed clinical isolates. Overall, amino acid sequences of clinical isolates were highly conserved compared to the prototype (99.2 to 100%), but a synonymous/non synonymous ratio (S/N) of 2.36 in E3 CR1 beta suggested positive selection. Conclusion: Unique sequence features of HAdV-A31 may enhance its ability to escape the host's immune surveillance and may facilitate a promiscuous tropism for various tissues. Moderate evolution of clinical isolates did not indicate the emergence of new HAdV-A31 subtypes in the recent years.