Design and validation of a high-throughput assay to detect codon 146 polymorphisms in the caprine prion protein gene.
In sheep, scrapie susceptibility is so strongly associated with single nucleotide polymorphisms (SNPs) in the gene encoding the prion protein (PrP) that this linkage constitutes the basis for selective breeding strategies directed toward controlling the disease. For goats, in contrast, the association between scrapie susceptibility/resistance and variations in the PrP gene is far weaker, with only a few identified SNPs showing an influence on scrapie susceptibility. A recent survey of PrP genotypes in Cypriot goats, however, revealed the existence of a robust association between polymorphisms at codon 146 of the caprine PrP gene and resistance/susceptibility to natural scrapie. Here we describe here a high-throughput assay, based on homogeneous MassExtend technology coupled with mass spectrometry, for genotyping codon 146 of the caprine PrP gene. Our results demonstrate that this assay exhibits high accuracy, reproducibility, and repeatability, thereby making it suitable for large-scale SNP genotyping, as required for scrapie surveillance programs.
Arrizubieta
María J
MJ
Institute of Agrobiotechnology, Center of Research and Technology Hellas, Thermi, Thessaloniki, Greece.
Kanata
Eirini
E
Keklikoglou
Ioanna
I
Papasavva-Stylianou
Penelope
P
Toumazos
Pavlos
P
Panagiotidis
Cynthia H
CH
Sklaviadis
Theodoros
T
D000818:Animals; D003062:Codon; D003535:Cyprus; D004198:Disease Susceptibility / Q000662:veterinary; D005820:Genetic Testing; D005838:Genotype; D015511:Goat Diseases / Q000235:genetics; D006041:Goats / Q000235:genetics*; D046208:Microchip Analytical Procedures / Q000379:methods / Q000662:veterinary*; D011110:Polymorphism, Genetic; D011328:Prions / Q000235:genetics*; D012608:Scrapie / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D003062:Codon; D011328:Prions