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Journal of Virology, September 2007, p. 9605-9608, Vol. 81, No. 17
0022-538X/07/$08.00+0     doi:10.1128/JVI.00635-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Efficient In Vitro Amplification of Chronic Wasting Disease PrPRES{triangledown}

Timothy D. Kurt,1 Matthew R. Perrott,1 Carol J. Wilusz,1 Jeffrey Wilusz,1 Surachai Supattapone,2 Glenn C. Telling,3 Mark D. Zabel,1 and Edward A. Hoover1*

Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado,1 Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire,2 Department of Molecular Biology and Genetics, University of Kentucky, Lexington, Kentucky3

Received 25 March 2007/ Accepted 30 May 2007

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 x 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.

* Corresponding author. Mailing address: Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523. Phone: (970) 491-7587. Fax: (970) 491-0523. E-mail: edward.hoover{at}colostate.edu

{triangledown} Published ahead of print on 6 June 2007.

Journal of Virology, September 2007, p. 9605-9608, Vol. 81, No. 17
0022-538X/07/$08.00+0     doi:10.1128/JVI.00635-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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