Large-scale PRNP genotyping of small ruminants using an automated high-throughput MALDI-TOF MS assay.

Content Provider	Europe PMC
Author	Migliore, Sergio Bivona, Maurizio Gagliostro, Enrico Buttitta, Onofrio Lo Mascolo, Francesca Macrì, Daniele
Copyright Year	2019
Description	Scrapie resistance or susceptibility in sheep is associated with single nucleotide polymorphisms (SNPs) at codons 136, 154, and 171 of the prion protein gene (PRNP). In addition, phenylalanine mutation at codon 141 has been recognized as a risk factor for atypical scrapie. In contrast, K222, D146, and S146 alleles confer genetic resistance to classical scrapie in goats. High-throughput genotyping technologies would provide significant benefits in scrapie eradication plans. The ability to resolve oligonucleotides varying in mass by less than a single nucleotide makes MALDI-TOF mass spectrometry (MS) a suitable platform for PRNP genotyping. We evaluated the commercial Myriapod scrapie kit (Diatech Pharmacogenetics), associated with a highly automated processing platform incorporating MALDI-TOF MS technology, to detect SNPs at codons 136, 154, 171, 141, and 222 of small ruminant PRNP. The Myriapod scrapie kit was accredited according to UNI CEI EN ISO/IEC 17025. We present the genotyping results of 10,960 sheep in Sicily and 1,822 goats in Sicily and Calabria (southern Italy) tested during 2017. We found a high frequency (43.9%) of the protective ARR allele in sheep and a promising 12.3% of the resistant K222 variant in goats. This efficient and high-throughput method is suitable for extensive PRNP genotyping, as requested in the European scrapie eradication plan.
Abstract	Scrapie resistance or susceptibility in sheep is associated with singlenucleotide polymorphisms (SNPs) at codons 136, 154, and 171 of the prion proteingene (PRNP). In addition, phenylalanine mutation at codon 141has been recognized as a risk factor for atypical scrapie. In contrast, K222,D146, and S146 alleles confer genetic resistance to classical scrapie in goats.High-throughput genotyping technologies would provide significant benefits inscrapie eradication plans. The ability to resolve oligonucleotides varying inmass by less than a single nucleotide makes MALDI-TOF mass spectrometry (MS) asuitable platform for PRNP genotyping. We evaluated thecommercial Myriapod scrapie kit (Diatech Pharmacogenetics), associated with ahighly automated processing platform incorporating MALDI-TOF MS technology, todetect SNPs at codons 136, 154, 171, 141, and 222 of small ruminant PRNP. TheMyriapod scrapie kit was accredited according to UNI CEI EN ISO/IEC 17025. Wepresent the genotyping results of 10,960 sheep in Sicily and 1,822 goats inSicily and Calabria (southern Italy) tested during 2017. We found a highfrequency (43.9%) of the protective ARR allele in sheep and a promising 12.3% ofthe resistant K222 variant in goats. This efficient and high-throughput methodis suitable for extensive PRNP genotyping, as requested in theEuropean scrapie eradication plan.
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ISSN	10406387
Volume Number	31
DOI	10.1177/1040638719859043
PubMed Central reference number	PMC6857025
Issue Number	4
PubMed reference number	31238800
Journal	Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc [J Vet Diagn Invest]
e-ISSN	19434936
Language	English
Publisher	SAGE Publications
Publisher Date	2019-06-25
Publisher Place	Sage CA: Los Angeles, CA
Access Restriction	Open
Rights License	© 2019 The Author(s)
Subject Keyword	MALDI-TOF MS PRNP genotyping scrapie single nucleotide polymorphisms small ruminants
Content Type	Text
Resource Type	Article
Subject	Veterinary