Efficient in vitro amplification of chronic wasting disease PrPRES.
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.
Kurt
Timothy D
TD
Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
Perrott
Matthew R
MR
Wilusz
Carol J
CJ
Wilusz
Jeffrey
J
Supattapone
Surachai
S
Telling
Glenn C
GC
Zabel
Mark D
MD
Hoover
Edward A
EA
D000818:Animals; D001921:Brain / Q000473:pathology*; D003670:Deer; D005289:Ferrets; D051379:Mice; D008822:Mice, Transgenic; D011328:Prions / Q000302:isolation & purification / Q000378:metabolism*; D034081:Wasting Disease, Chronic / Q000175:diagnosis*
D011328:Prions