Prion protein gene polymorphisms in healthy and scrapie-affected sheep in Greece.

A total of 216 local crossbred sheep from 16 scrapie-affected Greek flocks and 210 purebred sheep of the milk breeds Chios and Karagouniko from healthy flocks were analysed for scrapie-linked polymorphisms in the prion protein (PrP) gene. Of the 216 sheep in this case-control study, 96 sheep were clinical cases, 25 subclinical cases (asymptomatic at the moment of euthanasia but positive by histopathology and/or ELISA detecting proteinase-resistant PrP) and 95 healthy controls (negative by all evaluations). Polymorphisms at codons 136, 154 and 171 were determined by denaturing gradient gel electrophoresis, followed by RFLP and sequencing. Scrapie, both clinical and subclinical, was associated with the genotypes ARQ/ARQ (88 of 110 sheep of that genotype), ARQ/TRQ (9 of 13), ARQ/AHQ (15 of 38) and VRQ/VRQ (9 of 17). Histopathological lesions were more severe in the clinical cases. Genotypes ARQ/ARR (26 sheep), ARQ/ARK (seven sheep), AHQ/ARR (one sheep), ARH/ARH (one sheep) and ARR/ARH (three sheep) were detected exclusively in healthy control sheep. In the purebred survey, four genotypes were present in the Chios sheep (ARQ/ARQ, ARQ/TRQ, ARQ/AHQ and ARQ/ARR) and four in the Karagouniko sheep (ARQ/ARQ, ARQ/AHQ, ARQ/ARR and ARQ/ARH).

Prion protein gene polymorphisms in natural goat scrapie.

A total of 51 goats, including seven clinical cases, from the first herd in Greece reported to have scrapie was examined to discern an association between scrapie susceptibility and polymorphisms of the gene encoding the prion protein (PrP). Each animal was evaluated for clinical signs of the disease, histopathological lesions associated with scrapie, the presence of detectable protease-resistant PrP in the brain and PrP genotype. Eleven different PrP genotypes encoding at least five unique predicted mature PrP amino acid sequences were found. These genotypes included the amino acid polymorphisms at codons 143 (H-->R) and 240 (S-->P) and 'silent' nucleotide alterations at codons 42 (a-->g) and 138 (c-->t). Additionally, novel caprine amino acid polymorphisms were detected at codons 21 (V-->A), 23 (L-->P), 49 (G-->S), 154 (R-->H), 168 (P-->Q) and 220 (Q-->H) and new silent mutations were found at codons 107 (g-->a) and 207 (g-->a). The following variants were found in scrapie-affected goats: VV(21), LL(23), GG(49,) SS(49), HH(143), HR(143), RR(154), PP(168), PP(240), SP(240) and SS(240). All scrapie-affected animals carried the HH(143)RR(154) genotype, with the exception of two goats (HR(143)), both of which had detectable protease-resistant PrP but showed no clinical signs or histopathological lesions characteristic of scrapie.