FIGURE 3.

PMCA detection of sporadic and variant CJD prions in suspension or bound to steel wires using BacV-derived human PrP substrate. Serial dilutions of humanized sporadic CJD subtypes and variant CJD (vCJD) prions passaged into human PrP mice [tg650 line (Met129 allele) or BacV line (Val129 allele)] were used in suspension (A,B) or bound to steel wires (C,D). They were mixed with uninfected BacV brain homogenate and submitted to the mb-PMCA for one round (A,B) or two rounds (C,D) of 96 cycles of sonication/incubation for 48 h. The PMCA products were analyzed for PrP^Sc content by immunoblot. (A,C) Summary of the results obtained. The red boxes indicate the presence of PrP^Sc in PMCA products, the ratio in each box corresponds to the number of PrP^Sc-positive amplification products over the number of PMCA replicates performed. (B,D) Representative immunoblots (Sha31 anti-PrP antibody). MM, molecular weight markers.