NR AUQJ
AU Schönenbrücher,H.; Abdulmawjood,A.; Bülte,M.
TI Detection of central nervous system tissues in meat products: Validation studies of a Real Time-PCR based detection system including a ring trial approval
QU TSE-Forum, 6. Kongress - Nationale TSE-Forschungsplattform, Greifswald 26.6.-28.6.2006, Vortrag V-02
PT Konferenz-Vortrag
AB
In previous studies we presented the development and the validation of a quantitative and species-specific real-time RT-PCR to identify BSE risk material in meat and meat products. This was based on an mRNA assay that used bovine, ovine, and caprine glial fibrillary acidic protein (GFAP) encoding gene sequences as a marker.
Furthermore a national ring trial was conducted. As a prerequisite the 12 German laboratories applied real-time PCR Thermocycler models provided by Applied Biosystems. All the required laboratory reagents were prepared and shipped to the participants by our institute. For each sample, the mRNA was isolated from cooked sausages prepared according to the German traditional recipe and transcribed into cDNA before. The qualitative results obtained from the collaborative trial gave a diagnostic specificity of 100% for all negative samples including swine brain, bovine DNA and sausages without addition of CNS tissues. The diagnostic sensitivity for the positive samples was calculated with a value of 96.43%. In addition the accordance (repeatability) was 95.37%. The concordance (reproducibility) was 92.85%, calculated from all positive inoculation levels.
The quantitative real-time RT-PCR detection of GFAP mRNA appears to be useful as a routine diagnostic test for the detection of illegal use of CNS tissues in meat and meat products. The stability of the specific region of GFAP-mRNA allows the detection of CNS tissues also after meat processing steps. The use of organ and species specific subunits of mRNA might be a promising approach also for the detection of other banned tissues.
This work is a part of a project founded by the Federal Ministry of Consumers Protection, Food and Agriculture (AZ.: 01HS022/1). For further information please visit: http://hrzntweb-v1.hrz.uni-giessen.de/forschungsbericht/layout/einzel95.cfm?FB=10&Institut=2200&lfd_Nr=50p.
AD H. Schönenbrücher, Institute of Veterinary Food Science, Faculty of Veterinary Medicine, Justus-Liebig-University Giessen, Germany
SP englisch
PO Deutschland
OR Tagungsband