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Volume 26, Number 1—January 2020
Research

Preclinical Detection of Prions in Blood of Nonhuman Primates Infected with Variant Creutzfeldt-Jakob Disease

Luis Concha-Marambio1, Marcelo A. Chacon, and Claudio SotoComments to Author 
Author affiliations: University of Texas, Houston, Texas, USA (L. Concha-Marambio, M.A. Chacon, C. Soto); Universidad de los Andes, Santiago, Chile (L. Concha-Marambio, C. Soto)

Main Article

Figure 4

PMCA optimization for detection of macaque-adapted vCJD (m-vCJD) prions in preclinical blood samples. A) Tenfold serial dilutions of m-vCJD BH were amplified by the regular PMCA substrate (right panel) and substrate supplemented with 100 μg/mL heparin (left panel). After completion of 2 PMCA rounds, samples were digested with 50 μg/mL of PK and analyzed by Western blot. B) PL and BC (500 μL) samples collected 1 month before disease onset from 3 m-vCJD infected macaques (M1, M2, and M3) were sark

Figure 4. PMCA optimization for detection of macaque-adapted vCJD (m-vCJD) prions in preclinical blood samples. A) Tenfold serial dilutions of m-vCJD BH were amplified by the regular PMCA substrate (right panel) and substrate supplemented with 100 μg/mL heparin (left panel). After completion of 2 PMCA rounds, samples were digested with 50 μg/mL of PK and analyzed by Western blot. B) PL and BC (500 μL) samples collected 1 month before disease onset from 3 m-vCJD infected macaques (M1, M2, and M3) were sarkosyl precipitated and analyzed by 4 PMCA rounds using hep-substrate. Samples from noninfected macaques were analyzed as negative controls (–). N refers to transgenic mouse normal BH without proteinase K treatment, which was used as a migration control. BH, brain homogenate; PMCA, protein misfolding cyclic amplification; vCJD, variant Creutzfeldt-Jakob disease.

Main Article

1Current affiliation: Amprion, Inc., San Diego, California, USA.

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