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Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion
Authors:Atarashi Ryuichiro  Satoh Katsuya  Sano Kazunori  Fuse Takayuki  Yamaguchi Naohiro  Ishibashi Daisuke  Matsubara Takehiro  Nakagaki Takehiro  Yamanaka Hitoki  Shirabe Susumu  Yamada Masahito  Mizusawa Hidehiro  Kitamoto Tetsuyuki  Klug Genevieve  McGlade Amelia  Collins Steven J  Nishida Noriyuki
Institution:Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan. atarashi@nagasaki-u.ac.jp
Abstract:The development of technologies for the in vitro amplification of abnormal conformations of prion protein (PrP(Sc)) has generated the potential for sensitive detection of prions. Here we developed a new PrP(Sc) amplification assay, called real-time quaking-induced conversion (RT-QUIC), which allows the detection of ≥1 fg of PrP(Sc) in diluted Creutzfeldt-Jakob disease (CJD) brain homogenate. Moreover, we assessed the technique first in a series of Japanese subjects and then in a blind study of 30 cerebrospinal fluid specimens from Australia, which achieved greater than 80% sensitivity and 100% specificity. These findings indicate the promising enhanced diagnostic capacity of RT-QUIC in the antemortem evaluation of suspected CJD.
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