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• W2100051689 abstract "ABSTRACT 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 Creutzfeldt-Jakob disease." @default.
• W2100051689 created "2016-06-24" @default.
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• W2100051689 creator A5083992214 @default.
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• W2100051689 date "2014-02-28" @default.
• W2100051689 modified "2023-10-18" @default.
• W2100051689 title "Highly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification" @default.
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• W2100051689 doi "https://doi.org/10.1128/mbio.00829-13" @default.
• W2100051689 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3884057" @default.
• W2100051689 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/24381300" @default.
• W2100051689 hasPublicationYear "2014" @default.
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