Table 2. PrP^res detection results in PMCA reactions seeded with white blood cells (WBC) or buffy coat (BC) from Cynomologus macaques clinically affected with vCJD.

		1/10				1/50				1/100
		R2	R3	R4	R5	R2	R3	R4	R5	R2	R3	R4	R5
Macaque 6	WBC	−	+	+	+	−	+	+	+	−	+	+	+
	BC	−	−	−	−	−	−	+	+	−	−	+	+
Macaque 8	WBC	−	+	+	+	−	+	+	+	−	−	+	+
	BC	−	−	−	+	−	−	+	+	−	−	+	+

Cynomologus macaques were intravenously challenged with (i) blood from a vCJD affected macaque (macaque 6) or (ii) human vCJD brain homogenate (macaque 8). WBC and BC were prepared from the same blood sample (5 mL citrate dextrose anticoagulant) collected from the clinically affected primate (just prior euthanasia); macaques 6 and 8 were respectively euthanased at 38 months post inoculation (mpi) and 51 mpi and sampled at that time (see figure 4).

WBC and BC were homogenized and then diluted 1/10, 1/50 and 1/100 in PMCA buffer before seeding PMCA reactions. Brain from transgenic mice that expressed the ARQ variant of the ovine PrP was used as substrate. Samples were submitted to 5 successive rounds (R) of amplification. PrP^res detection was carried in each PMCA reaction by Western blot (WB) using Sha31 anti PrP antibody (epitope epitope: YEDRYYRE, amino acid 145–152).