Patent ID: 8216788

Claim:
A method for detecting protease resistant prion protein (PrP-res) in a sample comprising: (a) mixing the sample with purified recombinant protease-sensitive prion protein (rPrP-sen) to make a reaction mix, wherein the rPrP-sen comprises the amino acid sequence set forth as SEQ ID NO: 3; (b) performing an amplification reaction comprising: (i) incubating the reaction mix at about 37° C. to permit coaggregation of the rPrP-sen with the PrP-res that may be present in the reaction mix, and maintaining incubation conditions that promote coaggregation of the rPrP-sen with the PrP-res and result in a conversion of the rPrP-sen to the recombinant protease resistant prion protein initiated by the presence of prions (rPrP-res (Sc) ), which is initiated by the presence of PrP-res in the sample, while inhibiting development of spontaneously occurring recombinant prion protein (rPrP-res (spon) ); (ii) agitating aggregates formed during step (i), in shaking cycles, wherein each shaking cycle of the shaking cycles comprises a period of rest and a period of shaking, and wherein the period of rest is about 30 seconds in length and the period of shaking is about 30 seconds in length, wherein agitating is performed for about 48 hours in the absence of sonication; and (c) detecting rPrP-res (Sc) in the reaction mix after agitating for about 48 hours as above, wherein detection of rPrP-res (Sc) in the reaction mix indicates that PrP-res was present in the sample.