NR ALCI
AU Stack,M.J.; Aldrich,A.M.; Davis,L.A.
TI Comparison of detergent and protease enzyme combinations for the detection of scrapie-associated fibrils from the central nervous system of sheep naturally affected with scrapie
QU Journal of Comparative Pathology 1997 Feb; 116(2): 181-9
PT journal article
AB Standardized samples of tissue from the central nervous system of four sheep naturally affected with scrapie and from four healthy control sheep were subjected to a centrifugal extraction technique used to obtain scrapie-associated fibrils; the latter were then demonstrated by negative-contrast transmission electron microscopy. This regime was used to evaluate the fibril yield obtained from the 25 possible combinations of five different detergents and five different proteolytic enzymes. N-lauroylsarcosine detergent was found to be the most efficient detergent for all five enzymes, followed by sulphabetaine 3-14. Sodium dodecyl sulphate detergent was successful only in combination with a subtilisin Carlsberg enzyme. Octylglucoside and nonidet P40 detergents did not produce fibrils with any of the enzymes. Proteinase K was the least efficient of the five enzymes when used in combination with N-lauroylsarcosine; subtilisin Carlsberg, clostripain, pronase and trypsin enzymes all gave higher fibril yields. A combination of N-lauroylsarcosine detergent and subtilisin Carlsberg proteolytic enzyme gave the highest fibril yield.
MH Animal; Central Nervous System/*pathology; Comparative Study; Detergents/*diagnostic use; Drug Combinations; Endopeptidases/*diagnostic use; Female; Microscopy, Electron; Neurofibrils/*ultrastructure; Scrapie/*pathology; Sheep; Sheep Diseases/*pathology
AD Veterinary Laboratories Agency, Central Veterinary Laboratory, Weybridge, Surrey, UK
SP englisch
PO England