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
ABSTRACT
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.
snip...
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
TSS
posted by Terry S. Singeltary Sr. at
5:28 PM
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