NR AUSS
AU Schütz,E.; Urnovitz,H.B.; Iakoubov,L.; Beck,J.; Cervenakova,L.; Czub,S.; Graham,C.; Dudas,S.; Groschup,M.H.; Schulz-Schaeffer,W.J.; Young,A.; Brenig,B.
TI Circulating Nucleic Acid-based Disgnostics for Bovine and Human Transmissible Spongiform Encephalopathies
QU TSE-Forum, 6. Kongress - Nationale TSE-Forschungsplattform, Greifswald 26.6.-28.6.2006, Poster: Diagnostik DIA-08
PT Konferenz-Poster
AB Several ante mortem tests for transmissible spongiform encephalopathies (TSEs) are currently under development. A reliable test will have significant value in animal TSE eradication programs, and as a vital screening tool to halt the spread of TSEs through transfusion of blood products and components. One current research area to be considered for ante mortem testing is the use of circulating nucleic acids (CNAs) in a Nucleic Acid Test (NAT) format. CNAs are defined as DNA or RNA found in acellular fluids, e.g., serum or plasma. We have previously reported that specific CNAs can be found in both PrPres confirmed cows and a majority of their feeding cohorts but not unexposed cattle (Schütz, et al. CDLI 12:814, 2005). In an effort to further confirm the efficacy of this approach for TSE diagnostics, two experimental studies were conducted on cows inoculated either orally or intracerebrally with BSE infectious brain material. In the first study, twenty (20) Fleckvieh cattle were fed 100 g of PrPres positive brain stem macerate and monitored at defined time points. Blood samples for the investigations were taken either 30 (10 animals) or 32 (10 animals) months post-inoculation. CNAs were pre-amplified and analyzed as previously described. Data demonstrated that 19 of 20 animals expressed BSE-specific CNA sequences 30 or 32 months post-inoculation, whereas little reactivity was observed in a set of normal control cows. In a second study, calves were inoculated intracerebrally with 3 distinct isolates of BSE. Animals were tested at 3, 4 and 5 months post inoculation. There was a time dependent increase in BSE-specific CNAs in the serum of these animals, detected in 1 out of 3 animals after 3 months, in 3 out of 3 animals at 4 months, and in 4 out of 4 experimental animals 5 months post-inoculation To investigate the utility of such CNAs across species barriers, a set of primers was developed to reveal CJD-specific CNAs in human samples. Using these primers, 8 of 8 German CJD sera were reactive in the assay, while 90 random German blood donors were non-reactive. Additionally, 5 of 5 American CJD sera were found to be reactive, whilst normal blood donors showed no reaction. Combined, these data further support evidence of the presence of TSE-associated CNA sequences in the peripheral blood of BSE-infected cattle and humans incubating CJD. Future work will be focused on larger validation studies, which will include other medical and veterinary conditions, and a definition of the mechanism behind the generation of the disease-related nucleic acid sequences. The data point to CNA-based NAT approaches as potentially viable tools in the pursuit of sensitive, inexpensive ante mortem TSE screening diagnostics.
AD Ekkehard Schütz, Julia Beck, Bertram Brenig, Institute of Veterinary Medicine, University of Göttingen, Germany; Ekkehard Schütz, Howard B. Urnovitz , Leonid Iakoubov, Julia Beck, Chronix Biomedical GmbH, Göttingen, Germany; Larisa Cervenáková, The American Red Cross, Rockville, MD, USA; Stefanie Czub, Canadian Food Inspection Agency, Winnipeg, Canada; Catherine Graham, Sandor Dudas, Canadian Food Inspection Agency, Lethbridge, Canada; Martin H. Groschup, Friedrich-Loeffler-Institut, German Federal Research Institute for Animal Health, Greifswald - Insel Riems, Germany; Walter J. Schulz-Schaeffer, Dept. of Neuropathology, University of Göttingen, Germany, Department of Veterinary Sciences, USA; Alan Young, South Dakota State University, Brookings, SD, USA
SP englisch
PO Deutschland
OR Tagungsband