Protein misfolding cyclic amplification as a rapid test for assessment of prion inactivation

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• 作者：Yuichi Murayama ; Miyako Yoshioka ; Hiroko Horii ; Masuhiro Takata ; Takashi Yokoyama ; Takashi Sudo ; Koichi Sato ; Morikazu Shinagawa and Shirou Mohri
• 关键词：Prion inactivation ; Incineration ; Autoclaving ; Bioassay ; Protein misfolding cyclic amplification
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2006
• 出版时间：22 September 2006
• 年：2006
• 卷：348
• 期：2
• 页码：758-762
• 全文大小：158 K

文摘

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 °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.