title: Protein misfolding cyclic amplification as a rapid test for assessment of prion inactivation.
abstract: Abnormal isoform of prion proteins (PrP(Sc)), which are infectious agents associated with prion diseases, retain infectivity after undergoing routine sterilization processes. A sensitive method to detect the infectivity is a bioassay, and it has been used for assessing prion inactivation. However, the result is obtained after several hundred days. Here, protein misfolding cyclic amplification (PMCA) in which PrP(Sc) can be amplified in vitro was applied for assessing prion inactivation by dry heating and autoclaving. Scrapie-infected hamster brains were inactivated under various conditions, and residual infectivity and PrP(Sc) were detected by the bioassay and PMCA, respectively. The PMCA results were in good agreement with those of the bioassay. In samples autoclaved at temperatures below 150 degrees C, while infected mice died in the bioassay, protease-resistant PrP (PrP(res)) signals were detected in the second or third round of PMCA. Three rounds of PMCA require only 6 days, which means that the PMCA method is much faster than the bioassay.
authors.affiliation: Prion Disease Research Center, National Institute of Animal Health, Ibaraki, Japan. ymura@affrc.go.jp
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Murayama
authors.forename: Miyako
authors.initials: M
authors.lastname: Yoshioka
authors.forename: Hiroko
authors.initials: H
authors.lastname: Horii
authors.forename: Masuhiro
authors.initials: M
authors.lastname: Takata
authors.forename: Takashi
authors.initials: T
authors.lastname: Yokoyama
authors.forename: Takashi
authors.initials: T
authors.lastname: Sudo
authors.forename: Koichi
authors.initials: K
authors.lastname: Sato
authors.forename: Morikazu
authors.initials: M
authors.lastname: Shinagawa
authors.forename: Shirou
authors.initials: S
authors.lastname: Mohri
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000379:methods; D006224:Cricetinae; D051379:Mice; D018620:PrPSc Proteins / Q000737:chemistry / Q000472:pathogenicity*; D017510:Protein Folding
chemical_list: D018620:PrPSc Proteins