NR AOUM
AU Kashkevich,X.; Humeny,A.; Becker,C.M.; Schiebel,K.
TI Prnp genotyping by MALDI-TOF-MS in control and BSE affected animals
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - DG-43
PT Konferenz-Poster
AB
In man and sheep, DNA polymorphisms of the prion protein gene PRNP/Prnp have been shown to be involved in susceptibility/resistance to infections with PrPsc. In contrast, genetic risk factors affecting the bovine form of transmissible spongiform encephalopathies (BSE) are poorly understood. To address this question, a two-step approach was used to analyze potential genetic factors affecting susceptibility/resistance in BSE: Initially, DNA-sequencing was performed to detect genetic variations in different bovine breeds. In the second step, the high-throughput technique MALDI-TOF-MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry) was established to genotype a high number of control and BSE-affected animals. For analysis of potential genetic risk factor, these protocols will be suitable for large scale genetic survey of cattle herds. The identification of potential susceptible genotypes will be helpful for breeding resistant animals, with MALDI-TOF-MS offering an efficient screening tool.
In detail, comparative sequencing of individuals of 6 different bovine breeds (Holstein Friesian = Schwarzbunt, Red Holstein = Rotbunt, Braunvieh = (German) Swiss Brown, Simmental = Fleckvieh, Gelbvieh and Limousin) resulted in the isolation of 24 polymorphisms predominantly single nucleotide polymorphisms (SNPs) within the prion protein gene. MALDI-TOF-MS based genotyping assays using primer extension reactions were established for these sequence variations and tested for automated genotyping. Up to now, >2000 genotypes in breeding bulls and >200 genotypes of DNAs from German BSE positive tested animals were obtained. Our initial genotyping results indicate a specific predominant genetic variation in the prion protein gene of BSE positive tested animals. Ongoing analysis using larger populations will be performed to clarify the existence of this and other genetic risk factors.
AD X. Kashkevich, A. Humeny, C.-M. Becker, K. Schiebel, Universität Erlangen-Nürnberg, Germany
SP englisch
PO Deutschland