Efficient in vitro amplification of chronic wasting disease PrPRES.

Abstract

Chronic wasting disease (CWD) of cervids is associated with conversion of the normal cervid prion protein, PrP(C), to a protease-resistant conformer, PrP(CWD). Here we report the use of both nondenaturing amplification and protein-misfolding cyclic amplification (PMCA) to amplify PrP(CWD) in vitro. Normal brains from deer, transgenic mice expressing cervid PrP(C) [Tg(cerPrP)1536 mice], and ferrets supported amplification. PMCA using normal Tg(cerPrP)1536 brains as the PrP(C) substrate produced >6.5 x 10(9)-fold amplification after six rounds. Highly efficient in vitro amplification of PrP(CWD) is a significant step toward detection of PrP(CWD) in the body fluids or excreta of CWD-susceptible species.