@article{BittorfBergmannMerlinetal.2020,
  author    = {Bittorf, Patrick and Bergmann, Thorsten and Merlin, Simone and Olgasi, Chistina and Pullig, Oliver and Sanzenbacher, Ralf and Zierau, Martin and Walles, Heike and Follenzi, Antonia and Braspenning, Joris},
  title     = {Regulatory-Compliant Validation of a Highly Sensitive qPCR for Biodistribution Assessment of Hemophilia A Patient Cells},
  series = {Molecular Therapy - Methods \& Clinical Development},
  volume    = {18},
  journal   = {Molecular Therapy - Methods \& Clinical Development},
  doi       = {10.1016/j.omtm.2020.05.029},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-230284},
  pages     = {176-188},
  year      = {2020},
  abstract  = {The investigation of the biodistribution profile of a cell-based medicinal product is a pivotal prerequisite to allow a factual benefit-risk assessment within the non-clinical to clinical translation in product development. Here, a qPCR-based method to determine the amount of human DNA in mouse DNA was validated according to the guidelines of the European Medicines Agency and the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. Furthermore, a preclinical worst-case scenario study was performed in which this method was applied to investigate the biodistribution of 2 x 10\(^6\) intravenously administered, genetically modified, blood outgrowth endothelial cells from hemophilia A patients after 24 h and 7 days. The validation of the qPCR method demonstrated high accuracy, precision, and linearity for the concentration interval of 1:1 x 10\(^3\) to 1:1 x 10\(^6\) human to mouse DNA. The application of this method in the biodistribution study resulted in the detection of human genomes in four out of the eight investigated organs after 24 h. After 7 days, no human DNA was detected in the eight organs analyzed. This biodistribution study provides mandatory data on the toxicokinetic safety profile of an actual candidate cell-based medicinal product. The extensive evaluation of the required validation parameters confirms the applicability of the qPCR method for non-clinical biodistribution studies.},
  language  = {en}
}
@article{PaholcsekFidlerKonyaetal.2015,
  author    = {Paholcsek, Melinda and Fidler, Gabor and Konya, Jozsef and Rejto, Laszlo and Mehes, Gabor and Bukta, Evelin and Loeffler, Juergen and Biro, Sandor},
  title     = {Combining standard clinical methods with PCR showed improved diagnosis of invasive pulmonary aspergillosis in patients with hematological malignancies and prolonged neutropenia},
  series = {BMC Infectious Diseases},
  volume    = {15},
  journal   = {BMC Infectious Diseases},
  number    = {251},
  doi       = {10.1186/s12879-015-0995-8},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-151607},
  year      = {2015},
  abstract  = {Background: We assessed the diagnostic value of standard clinical methods and combined biomarker testing (galactomannan assay and polymerase chain reaction screening) in a prospective case-control study to detect invasive pulmonary aspergillosis in patients with hematological malignancies and prolonged neutropenia. Methods: In this observational study 162 biomarker analyses were performed on samples from 27 febrile neutropenic episodes. Sera were successively screened for galactomannan antigen and for Aspergillus fumigatus specific nucleic acid targets. Furthermore thoracic computed tomography scanning was performed along with bronchoscopy with lavage when clinically indicated. Patients were retrospectively stratified to define a case-group with "proven" or "probable" invasive pulmonary aspergillosis (25.93 \%) and a control-group of patients with no evidence for of invasive pulmonary aspergillosis (74.07 \%). In 44.44 \% of episodes fever ceased in response to antibiotic treatment (group II). Empirical antifungal therapy was administered for episodes with persistent or relapsing fever (group I). 48.15 \% of patients died during the study period. Postmortem histology was pursued in 53.85 \% of fatalities. Results: Concordant negative galactomannan and computed tomography supported by a polymerase chain reaction assay were shown to have the highest discriminatory power to exclude invasive pulmonary aspergillosis. Bronchoalveolar lavage was performed in 6 cases of invasive pulmonary aspergillosis and in 15 controls. Although bronchoalveolar lavage proved negative in 93 \% of controls it did not detect IPA in 86 \% of the cases. Remarkably post mortem histology convincingly supported the presence of Aspergillus hyphae in lung tissue from a single case which had consecutive positive polymerase chain reaction assay results but was misdiagnosed by both computed tomography and consistently negative galactomannan assay results. For the galactomannan enzyme-immunoassay the diagnostic odds ratio was 15.33 and for the polymerase chain reaction assay it was 28.67. According to Cohen's kappa our in-house polymerase chain reaction method showed a fair agreement with the galactomannan immunoassay. Combined analysis of the results from the Aspergillus galactomannan enzyme immunoassay together with those generated by our polymerase chain reaction assay led to no misdiagnoses in the control group. Conclusion: The data from this pilot-study demonstrate that the consideration of standard clinical methods combined with biomarker testing improves the capacity to make early and more accurate diagnostic decisions.},
  language  = {en}
}