FIGURE 6.
Inhibition of distinct prion strains/isolates with rVRQ. All samples were amplified in duplicate by sPMCA for 5 rounds. Classical scrapie (genotypes VRQ/ARQ and VRQ/VRQ, isolates PG1361/05 and PG1563/02, respectively), transgenic-mouse passaged classical scrapie (2 isolates each of G338 and Apl338/Apl338ii passaged in mice with genotype VRQ) and experimental ovine BSE (2 isolates, both ARQ/ARQ) were amplified using a VRQ/VRQ PrPC substrate. Experimental CH1641 scrapie was amplified using an AHQ/AHQ PrPC substrate. Two bovine BSE samples were amplified using a bovine PrPC substrate. Representative western blots of PMCA reaction products amplified in the presence (+) or absence (−) of 400 nM rVRQ are shown for each TSE type (A). 6.7 µL of PK digested PMCA reaction was analyzed by western blotting using monoclonal antibody SHa31. Following densitometry, signals were expressed as the percent inhibition, calculated using the no inhibition controls as 100% amplification (B). The data in (B) is collated from 2 separate experiments. Molecular mass markers are indicated.