Highly infectious prions generated by a single round of microplate-based protein misfolding cyclic amplification.
mBio. 2013-12-31; 5(1):
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1. MBio. 2013 Dec 31;5(1):e00829-13. doi: 10.1128/mBio.00829-13.
Highly infectious prions generated by a single round of microplate-based protein
misfolding cyclic amplification.
Moudjou M, Sibille P, Fichet G, Reine F, Chapuis J, Herzog L, Jaumain E,
Laferrière F, Richard CA, Laude H, Andréoletti O, Rezaei H, Béringue V.
Measurements of the presence of prions in biological tissues or fluids rely more
and more on cell-free assays. Although protein misfolding cyclic amplification
(PMCA) has emerged as a valuable, sensitive tool, it is currently hampered by its
lack of robustness and rapidity for high-throughput purposes. Here, we made a
number of improvements making it possible to amplify the maximum levels of
scrapie prions in a single 48-h round and in a microplate format. The
amplification rates and the infectious titer of the PMCA-formed prions appeared
similar to those derived from the in vivo laboratory bioassays. This enhanced
technique also amplified efficiently prions from different species, including
those responsible for human variant Creutzfeldt-Jakob disease. This new format
should help in developing ultrasensitive, high-throughput prion assays for
cognitive, diagnostic, and therapeutic applications. IMPORTANCE The method
developed here allows large-scale, fast, and reliable cell-free amplification of
subinfectious levels of prions from different species. The sensitivity and
rapidity achieved approach or equal those of other recently developed
prion-seeded conversion assays. Our simplified assay may be amenable to
high-throughput, automated purposes and serve in a complementary manner with
other recently developed assays for urgently needed antemortem diagnostic tests,
by using bodily fluids containing small amounts of prion infectivity. Such a
combination of assays is of paramount importance to reduce the transfusion risk
in the human population and to identify asymptomatic carriers of variant
PMID: 24381300 [Indexed for MEDLINE]