Highly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification

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-throughp...

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Autores principales:	Mohammed Moudjou, Pierre Sibille, Guillaume Fichet, Fabienne Reine, Jérôme Chapuis, Laetitia Herzog, Emilie Jaumain, Florent Laferrière, Charles-Adrien Richard, Hubert Laude, Olivier Andréoletti, Human Rezaei, Vincent Béringue
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/020b9d4b8435459aa95ff2b0fda596ee
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spelling	oai:doaj.org-article:020b9d4b8435459aa95ff2b0fda596ee2021-11-15T15:45:11ZHighly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification10.1128/mBio.00829-132150-7511https://doaj.org/article/020b9d4b8435459aa95ff2b0fda596ee2014-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00829-13https://doaj.org/toc/2150-7511ABSTRACT 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.Mohammed MoudjouPierre SibilleGuillaume FichetFabienne ReineJérôme ChapuisLaetitia HerzogEmilie JaumainFlorent LaferrièreCharles-Adrien RichardHubert LaudeOlivier AndréolettiHuman RezaeiVincent BéringueAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 1 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Mohammed Moudjou Pierre Sibille Guillaume Fichet Fabienne Reine Jérôme Chapuis Laetitia Herzog Emilie Jaumain Florent Laferrière Charles-Adrien Richard Hubert Laude Olivier Andréoletti Human Rezaei Vincent Béringue Highly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification
description	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.
format	article
author	Mohammed Moudjou Pierre Sibille Guillaume Fichet Fabienne Reine Jérôme Chapuis Laetitia Herzog Emilie Jaumain Florent Laferrière Charles-Adrien Richard Hubert Laude Olivier Andréoletti Human Rezaei Vincent Béringue
author_facet	Mohammed Moudjou Pierre Sibille Guillaume Fichet Fabienne Reine Jérôme Chapuis Laetitia Herzog Emilie Jaumain Florent Laferrière Charles-Adrien Richard Hubert Laude Olivier Andréoletti Human Rezaei Vincent Béringue
author_sort	Mohammed Moudjou
title	Highly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification
title_short	Highly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification
title_full	Highly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification
title_fullStr	Highly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification
title_full_unstemmed	Highly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification
title_sort	highly infectious prions generated by a single round of microplate-based protein misfolding cyclic amplification
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/020b9d4b8435459aa95ff2b0fda596ee
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Highly Infectious Prions Generated by a Single Round of Microplate-Based Protein Misfolding Cyclic Amplification

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