IN VITRO DETECTION OF PRIONS IN BLOOD
A method of screening a blood sample for the presence of prions. The method includes the steps of collecting the blood sample in heparin, contacting the sample with a solution comprising recombinant prion protein (rPrP) and Thioflavin T (ThT), and measuring the resulting ThT fluorescence in the sample. The method can further include the step of freezing and thawing the sample prior to contacting the sample with a solution comprising recombinant prion protein (rPrP) and Thioflavin T (ThT). The method can also include the step of precipitating the prions in sodium phosphotungstic acid (NaPTA) prior to contacting the sample with a solution comprising recombinant prion protein (rPrP) and Thioflavin T (ThT).
1 . A method of screening a blood sample for the presence of prions comprising the steps of:
collecting the blood sample in heparin;
contacting the sample with a solution comprising recombinant prion protein (rPrP) and Thioflavin T (ThT); and
measuring the resulting ThT fluorescence in the sample.
2 . The method according to claim 1 wherein about 200 units/ml of heparin are added to the sample.
3 . The method according to claim 1 further comprising the step of freezing and thawing the sample prior to contacting the sample with a solution comprising recombinant prion protein (rPrP) and Thioflavin T (ThT).
4 . The method according to claim 3 wherein aliquots of whole blood were frozen at about −80° C. for about 30 minutes and subsequently thawed at about 22° C. for about 60 minutes.
5 . The method according to claim 3 wherein the samples are subjected to at least four (4) freeze-thaw cycles.
6 . The method according to claim 3 further comprising the step of homogenizing the sample after the freezing and thawing steps.
7 . The method according to claim 1 further comprising the step of precipitating the prions in sodium phosphotungstic acid (NaPTA) prior to contacting the sample with a solution comprising recombinant prion protein (rPrP) and Thioflavin T (ThT).