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Ultrasensitive detection of scrapie prion protein derived from ARQ and AHQ homozygote sheep by interspecies in vitro amplification
Ist Teil von
Microbiology and immunology, 2012-08, Vol.56 (8), p.541-547
Ort / Verlag
Melbourne, Australia: Blackwell Publishing Asia
Erscheinungsjahr
2012
Link zum Volltext
Quelle
Alma/SFX Local Collection
Beschreibungen/Notizen
ABSTRACT
Prions, infectious agents causing TSEs, are composed primarily of the pathogenic form (PrPSc) of the PrPC. The susceptibility of sheep to scrapie is determined by polymorphisms in the coding region of the PRNP, mainly at codons 136, 154, and 171. The efficiency of in vitro amplification of sheep PrPSc seems to be linked also to the PrP genotype. PrPSc derived from sheep with V136R154Q171‐associated genotypes can be amplified efficiently by PMCA in the presence of additional polyanion such as poly A, but there are no reports that cite ultrasensitive detection of PrPSc derived from sheep of other PrP genotypes. We report here that sheep PrPSc derived from ARQ and AHQ homozygotes was amplified efficiently by serial PMCA using mouse brain homogenate as PrPC substrate. ARQ/ARQ PrPSc was detected in infected brain homogenates diluted up to 10−10 after five rounds of amplification, and AHQ/AHQ PrPSc was detected in samples diluted up to 10−8 after four rounds of amplification. On the other hand, amplification of PrPSc from VRQ/ARQ sheep seemed to be less efficient under the experimental conditions used. The interspecies PMCA developed in this study may be useful in the detailed analysis of PrPSc distribution in classical scrapie‐infected ARQ and AHQ homozygote sheep.