title: Rapid and Highly Sensitive Detection of Variant Creutzfeldt-Jakob Disease Abnormal Prion Protein on Steel Surfaces by Protein Misfolding Cyclic Amplification: Application to Prion Decontamination Studies.
abstract: The prevalence of variant Creutzfeldt-Jakob disease (vCJD) in the population remains uncertain, although it has been estimated that 1 in 2000 people in the United Kingdom are positive for abnormal prion protein (PrPTSE) by a recent survey of archived appendix tissues. The prominent lymphotropism of vCJD prions raises the possibility that some surgical procedures may be at risk of iatrogenic vCJD transmission in healthcare facilities. It is therefore vital that decontamination procedures applied to medical devices before their reprocessing are thoroughly validated. A current limitation is the lack of a rapid model permissive to human prions. Here, we developed a prion detection assay based on protein misfolding cyclic amplification (PMCA) technology combined with stainless-steel wire surfaces as carriers of prions (Surf-PMCA). This assay allowed the specific detection of minute quantities (10-8 brain dilution) of either human vCJD or ovine scrapie PrPTSE adsorbed onto a single steel wire, within a two week timeframe. Using Surf-PMCA we evaluated the performance of several reference and commercially available prion-specific decontamination procedures. Surprisingly, we found the efficiency of several marketed reagents to remove human vCJD PrPTSE was lower than expected. Overall, our results demonstrate that Surf-PMCA can be used as a rapid and ultrasensitive assay for the detection of human vCJD PrPTSE adsorbed onto a metallic surface, therefore facilitating the development and validation of decontamination procedures against human prions.
authors.affiliation: Laboratoire TransDiag, UMR 1058, Etablissement Français du Sang Pyrénées-Méditerranée, Montpellier, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Belondrade
authors.affiliation: Laboratoire TransDiag, UMR 1058, Etablissement Français du Sang Pyrénées-Méditerranée, Montpellier, France.
authors.forename: Simon
authors.initials: S
authors.lastname: Nicot
authors.affiliation: Institut National de la Recherche Agronomique, UR892, Virologie Immunologie Moléculaires, Jouy-en-Josas, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Béringue
authors.affiliation: Laboratoire TransDiag, UMR 1058, Etablissement Français du Sang Pyrénées-Méditerranée, Montpellier, France.
authors.forename: Joliette
authors.initials: J
authors.lastname: Coste
authors.affiliation: CHRU de Montpellier and Université de Montpellier, IRMB, INSERM U1183, Laboratoire de Biochimie Protéomique Clinique, Montpellier, France.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Lehmann
authors.affiliation: Laboratoire TransDiag, UMR 1058, Etablissement Français du Sang Pyrénées-Méditerranée, Montpellier, France.
authors.forename: Daisy
authors.initials: D
authors.lastname: Bougard
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D001921:Brain / Q000473:pathology; D007562:Creutzfeldt-Jakob Syndrome / Q000473:pathology / Q000517:prevention & control* / Q000635:transmission; D003428:Cross Infection / Q000517:prevention & control* / Q000635:transmission; D003666:Decontamination / Q000379:methods*; D004864:Equipment and Supplies; D006801:Humans; D057230:Limit of Detection; D051379:Mice; D018620:PrPSc Proteins / Q000378:metabolism; D011328:Prions / Q000378:metabolism*; D017510:Protein Folding; D012608:Scrapie / Q000378:metabolism; D013193:Stainless Steel; D013499:Surface Properties; D006113:United Kingdom
chemical_list: D018620:PrPSc Proteins; D011328:Prions; D013193:Stainless Steel