Muscle sample prepared for prion assay
View Patent ↗A method of preparing a sample of muscle tissue and of assaying the prepared sample to determine the presence of prions in the sample is disclosed. The muscle tissue is homogenized and mixed with a complexing agent which forms a complex with a higher specific gravity than PrP Sc , the complexing agent or other components of the homogenate. Gravity is then used (e.g. ultra centrifugation) to concentrate the complex and the concentrate is assayed to detect prions. The muscle tissue is preferably extracted from a muscle or group of muscles such as hind limb muscle which have a higher or more preferably the highest concentration of prions as compared to other muscle in the mammal.
1. A method of preparing a sample, comprising the steps of:
obtaining a sample of muscle tissue from a mammal;
homogenizing the sample thereby creating a homogenate;
contacting the homogenate with a complexing agent which binds PrP Sc in the homogenate forming a complex with a higher specific gravity as compared to the specific gravity of either PrP Sc or the complexing agent alone; and
concentrating the complex by the application of gravity.
2. The method of claim 1 , wherein the muscle tissue is from muscle having a higher concentration of prions than other muscle for the species of mammal from which the sample is obtained.
3. The method of claim 2 , wherein the sample is obtained from muscle having a higher concentration of prions as compared to any other muscle in the mammal from which the muscle is obtained.
4. The method of claim 1 , wherein the sample of muscle tissue is obtained from hind limb muscle of the mammal chosen from a cow, a sheep and a deer wherein the concentration by gravity is enhanced by centrifugation.
5. The method of claim 1 , wherein the complexing agent is chosen from a heteropoly acid and a salt thereof.
6. The method of claim 5 , wherein the complexing agent is sodium phosphotungstate.
7. The method of claim 1 , further comprising:
treating the homogenate under conditions and for a period of time so as to denature PrP C in the homogenate and so as to change the conformation of PrP Sc in the homogenate.