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Journal of Clinical Microbiology, September 2003, p. 4127-4133, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4127-4133.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Molecular Analysis of Cases of Italian Sheep Scrapie and Comparison with Cases of Bovine Spongiform Encephalopathy (BSE) and Experimental BSE in Sheep

Romolo Nonno,1* Elena Esposito,1 Gabriele Vaccari,1 Michela Conte,1 Stefano Marcon,1 Michele Di Bari,1 Ciriaco Ligios,2 Giovanni Di Guardo,3,{dagger} and Umberto Agrimi1

Laboratory of Veterinary Medicine, Istituto Superiore di Sanità,1 Istituto Zooprofilattico Sperimentale del Lazio e della Toscana, Rome,3 Istituto Zooprofilattico Sperimentale della Sardegna, Sassari, Italy2

Received 7 March 2003/ Returned for modification 11 May 2003/ Accepted 22 June 2003

Concerns have been raised about the possibility that the bovine spongiform encephalopathy (BSE) agent could have been transmitted to sheep populations via contaminated feedstuffs. The objective of our study was to investigate the suitability of molecular strain typing methods as a surveillance tool for studying scrapie strain variations and for differentiating PrPSc from sheep scrapie, BSE, and sheep BSE. We studied 38 Italian sheep scrapie cases from 13 outbreaks, along with a British scrapie case, an experimental ovine BSE, and 3 BSE cases, by analyzing the glycoform patterns and the apparent molecular masses of the nonglycosylated forms of semipurified, proteinase-treated PrPSc. Both criteria were able to clearly differentiate sheep scrapie from BSE and ovine experimental BSE. PrPSc from BSE and sheep BSE showed a higher glycoform ratio and a lower molecular mass of the nonglycosylated form compared to scrapie PrPSc. Scrapie cases displayed homogeneous PrPSc features regardless of breed, flock, and geographic origin. The glycoform patterns observed varied with the antibody used, but either a monoclonal antibody (MAb) (F99/97.6.1) or a polyclonal antibody (P7-7) was able to distinguish scrapie from BSE PrPSc. While more extensive surveys are needed to further corroborate these findings, our results suggest that large-scale molecular screening of sheep populations for BSE surveillance may be eventually possible.

* Corresponding author. Mailing address: Laboratory of Veterinary Medicine, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy. Phone: 39-06-49902854. Fax: 39-06-49387077. E-mail: romolo.nonno{at}iss.it.

{dagger} Present address: University of Teramo, Faculty of Veterinary Medicine, Teramo, Italy.

Journal of Clinical Microbiology, September 2003, p. 4127-4133, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4127-4133.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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