tailieunhanh - Báo cáo khoa học: Hyperefficient PrPSc amplification of mouse-adapted BSE and scrapie strain by protein misfolding cyclic amplification technique

Abnormal forms of prion protein (PrP Sc ) accumulate via structural conver-sion of normal PrP (PrP C ) in the progression of transmissible spongiform encephalopathy. Under cell-free conditions, the process can be efficiently replicated usingin vitro PrP Sc amplification methods, including protein mis-folding cyclic amplification. | ỊFEBS Journal Hyperefficient PrPSc amplification of mouse-adapted BSE and scrapie strain by protein misfolding cyclic amplification technique Aiko Fujihara Ryuichiro Atarashi Takayuki Fuse Kaori Ubagai Takehiro Nakagaki Naohiro Yamaguchi Daisuke Ishibashi Shigeru Katamine and Noriyuki Nishida Department of Molecular Microbiology and Immunology Nagasaki University Graduate Schoolof BiomedicalSciences Japan Keywords prion protein misfolding cyclic amplification sonication transmissible spongiform encephalopathy Correspondence R. Atarashi Department of Molecular Microbiology and Immunology Nagasaki University Graduate Schoolof Biomedical Sciences 1-12-4 Sakamoto Nagasaki 852-8523 Japan Fax 81 95 819 7060 Tel 81 95 819 7060 E-mail atarashi@ Received 19 February 2009 revised 11 March 2009 accepted 16 March 2009 doi Abnormal forms of prion protein PrPSc accumulate via structural conversion of normal PrP PrPC in the progression of transmissible spongiform encephalopathy. Under cell-free conditions the process can be efficiently replicated using in vitro PrPSc amplification methods including protein misfolding cyclic amplification. These methods enable ultrasensitive detection of PrPSc however there remain difficulties in utilizing them in practice. For example to date several rounds of protein misfolding cyclic amplification have been necessary to reach maximal sensitivity which not only take several weeks but also result in an increased risk of contamination. In this study we sought to further promote the rate of PrPSc amplification in the protein misfolding cyclic amplification technique using mouse transmissible spongiform encephalopathy models infected with either mouse-adapted bovine spongiform encephalopathy or mouse-adapted scrapie Chandler strain. Here we demonstrate that appropriate regulation of sonication dramatically accelerates PrPSc amplification in both strains. In fact we reached maximum sensitivity allowing