Development and usefulness of new polymerase chain reaction-based tests for detection of different alleles at codons 136 and 171 of the ovine prion protein gene

Vilma Yuzbasiyan-Gurkan From the Department of Small Animal Clinical Sciences, College of Veterinary Medicine (Yuzbasiyan-Gurkan. Cao. Venta) and the Departments of Microbiology (Yuzbasiyan-Gurkan, Venta) and Veterinary Pathology (Krehbiel), Michigan State University, East Lansing, MI 48824-1314

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Janver D. Krehbiel From the Department of Small Animal Clinical Sciences, College of Veterinary Medicine (Yuzbasiyan-Gurkan. Cao. Venta) and the Departments of Microbiology (Yuzbasiyan-Gurkan, Venta) and Veterinary Pathology (Krehbiel), Michigan State University, East Lansing, MI 48824-1314

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Yueying Cao From the Department of Small Animal Clinical Sciences, College of Veterinary Medicine (Yuzbasiyan-Gurkan. Cao. Venta) and the Departments of Microbiology (Yuzbasiyan-Gurkan, Venta) and Veterinary Pathology (Krehbiel), Michigan State University, East Lansing, MI 48824-1314

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Patrick J. Venta From the Department of Small Animal Clinical Sciences, College of Veterinary Medicine (Yuzbasiyan-Gurkan. Cao. Venta) and the Departments of Microbiology (Yuzbasiyan-Gurkan, Venta) and Veterinary Pathology (Krehbiel), Michigan State University, East Lansing, MI 48824-1314

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Abstract

Objective

To develop new and improved tests to detect alleles at codons 136 and 171 of the ovine prion protein locus and to evaluate the frequency of these alleles.

Animals

159 Suffolk sheep belonging to 3 flocks.

Procedure

Polymerase chain reaction (PCR) analysis that contained diagnostic restriction site variation for each allele were developed for the relevant gene regions. Alleles were determined by analyzing DNA isolated from buccal swab specimens or blood samples.

Results

At codon 136, frequencies of the alanine and valine alleles were found to be 97 and 3%, respectively. At codon 171, frequencies of the glutamine, arginine, and histidine alleles were found to be 57, 41, and 2%, respectively.

Conclusions

Little variation was detected in codon 136, whereas noteworthy variation was found in codon 171; > 40% of the alleles at this locus coded for glutamine. Because the glutamine allele at codon 171 confers susceptibility to scrapie, reduction of its frequency is of importance to management of sheep flocks.

Clinical Relevance

Genotyping of sheep, using the tests reported here, should facilitate selective breeding programs designed to decrease the risk of scrapie. (Am J Vet Res 1999;60:884–887)

Abstract

Objective

To develop new and improved tests to detect alleles at codons 136 and 171 of the ovine prion protein locus and to evaluate the frequency of these alleles.

Animals

159 Suffolk sheep belonging to 3 flocks.

Procedure

Polymerase chain reaction (PCR) analysis that contained diagnostic restriction site variation for each allele were developed for the relevant gene regions. Alleles were determined by analyzing DNA isolated from buccal swab specimens or blood samples.

Results

At codon 136, frequencies of the alanine and valine alleles were found to be 97 and 3%, respectively. At codon 171, frequencies of the glutamine, arginine, and histidine alleles were found to be 57, 41, and 2%, respectively.

Conclusions

Little variation was detected in codon 136, whereas noteworthy variation was found in codon 171; > 40% of the alleles at this locus coded for glutamine. Because the glutamine allele at codon 171 confers susceptibility to scrapie, reduction of its frequency is of importance to management of sheep flocks.

Clinical Relevance

Genotyping of sheep, using the tests reported here, should facilitate selective breeding programs designed to decrease the risk of scrapie. (Am J Vet Res 1999;60:884–887)

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