Thursday, November 21, 2013

Efficient In Vitro Amplification of Chronic Wasting Disease PrPRES

Efficient In Vitro Amplification of Chronic Wasting Disease PrPRES▿


Timothy D. Kurt1, Matthew R. Perrott1, Carol J. Wilusz1, Jeffrey Wilusz1, Surachai Supattapone2, Glenn C. Telling3, Mark D. Zabel1, and Edward A. Hoover1,* + Author Affiliations


1Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado 2Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 3Department of Molecular Biology and Genetics, University of Kentucky, Lexington, Kentucky Next Section




Chronic wasting disease (CWD) of cervids is associated with conversion of the normal cervid prion protein, PrPC, to a protease-resistant conformer, PrPCWD. Here we report the use of both nondenaturing amplification and protein-misfolding cyclic amplification (PMCA) to amplify PrPCWD in vitro. Normal brains from deer, transgenic mice expressing cervid PrPC [Tg(cerPrP)1536 mice], and ferrets supported amplification. PMCA using normal Tg(cerPrP)1536 brains as the PrPC substrate produced >6.5 × 109-fold amplification after six rounds. Highly efficient in vitro amplification of PrPCWD is a significant step toward detection of PrPCWD in the body fluids or excreta of CWD-susceptible species.




 In summary, we report efficient amplification of PrPCWD by using brain substrates from Tg(cerPrP)1536 mice. The magnitude of PrPCWD conversion obtained by serial PMCA may make possible in vitro detection of PrPCWD in the body fluids and excreta of infected animals. These studies have been initiated.






Thursday, November 21, 2013


Assessing the susceptibility of transgenic mice over-expressing deer prion protein to bovine spongiform encephalopathy








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