NR AOWJ
AU Martucci,F.; Bozzetta,E.; Nappi,R.; Casalone,C.; Ferrari,A.; Mazza,M.; Riina,M.V.; Manzardo,E.; Caramelli,M.; Acutis,P.L.
TI Evaluation of BSE Rapid Tests for the Diagnosis of Transmissible Spongiform Encephalopathies in Sheep and Goats
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - DG-38
PT Konferenz-Poster
AB To improve TSE surveillance, according to EU Decision 999/2001/EC, rapid tests already adopted for BSE (Prionics, Abbott, Biorad) have to be applied to small ruminants. However, the tests have been evaluated only for BSE and no official data are available about their performances on scrapie. In our study we evaluated the accuracy of these methods for TSE diagnosis in Italian sheep and goats. 550 negative brainstems for TSE (500 sheep, 50 goats) were selected. The animals were of pure and mixed breeds, healthy slaughtered, older than 18 months. 38 positive brainstems for TSE from sheep of different breeds, found by active or passive surveillance, were collected; only 2 goats could be included. True negativity and positivity were confirmed by histology, immunohistochemistry and Western blot. PrP genotype of positive sheep was determined, with the following results: ARQ/ARQ (34), ARQ/AHQ (3), ARQ/VRQ (1). To warrant comparable results from the rapid tests, positive samples were subjected to an in-house pre-homogenisation protocol. Briefly 2 gr. of sample were cut in small pieces with two scalpels without any buffer, till the tissue appeared homogeneous. The obtained homogenate was split in three aliquots destined to the tests, according to the amount needed by the protocols. All three tests showed 100% of specificity and sensitivity for sheep (lower limit C.I. 95% = 99.0% and 95.2% respectively). Regarding goats, sensitivity was 100% (lower limit C.I. 95% = 91.1%), while paucity of samples didn't allow to estimate specificity. In conclusion, the three systems appeared to be reliable for active surveillance in sheep, while further studies are needed on goats. Besides, we set-up a new pre-homogenisation protocol that can be useful for future test evaluation, avoiding heterogeneity of the PrPsc in slices and its dilution caused by additional buffer. This research was founded by the Ministry of Health (IZSPLV001/2000).
AD F. Martucci, E. Bozzetta, R. Nappi, C. Casalone, A. Ferrari, M. Mazza, M.V. Riina, E. Manzardo, M. Caramelli, P.L. Acutis, CEA- Istituto Zooprofilattico di Torino
SP englisch
PO Deutschland