A direct assessment of human prion adhered to steel wire using real-time quaking-induced conversion.
Accidental transmission of prions during neurosurgery has been reported as a consequence of re-using contaminated surgical instruments. Several decontamination methods have been studied using the 263K-hamster prion; however, no studies have directly evaluated human prions. A newly developed in vitro amplification system, designated real-time quaking-induced conversion (RT-QuIC), has allowed the activity of abnormal prion proteins to be assessed within a few days. RT-QuIC using human recombinant prion protein (PrP) showed high sensitivity for prions as the detection limit of our assay was estimated as 0.12 fg of active prions. We applied this method to detect human prion activity on stainless steel wire. When we put wires contaminated with human Creutzfeldt-Jakob disease brain tissue directly into the test tube, typical PrP-amyloid formation was observed within 48 hours, and we could detect the activity of prions at 50% seeding dose on the wire from 10(2.8) to 10(5.8) SD50. Using this method, we also confirmed that the seeding activities on the wire were removed following treatment with NaOH. As seeding activity closely correlated with the infectivity of prions using the bioassay, this wire-QuIC assay will be useful for the direct evaluation of decontamination methods for human prions.
Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Tsuyoshi
T
Mori
Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Ryuichiro
R
Atarashi
Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Kana
K
Furukawa
Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Hanae
H
Takatsuki
Department of Locomotive Rehabilitation Science, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8523, Japan.
Katsuya
K
Satoh
Department of Physiology and Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Jonan-ku, Fukuoka 814-0180, Japan.
Kazunori
K
Sano
Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Takehiro
T
Nakagaki
Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Daisuke
D
Ishibashi
Kripton Co., Ltd., Dai 12 Daitetsu Bldg. 7F., 4-3-12 Yotsuya, Shinjuku-ku, Tokyo 160-0004, Japan.
Kazuko
K
Ichimiya
Kripton Co., Ltd., Dai 12 Daitetsu Bldg. 7F., 4-3-12 Yotsuya, Shinjuku-ku, Tokyo 160-0004, Japan.
Masahisa
M
Hamada
Kripton Co., Ltd., Dai 12 Daitetsu Bldg. 7F., 4-3-12 Yotsuya, Shinjuku-ku, Tokyo 160-0004, Japan.
Takehisa
T
Nakayama
Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Noriyuki
N
Nishida
D000682:Amyloid / Q000187:drug effects / Q000378:metabolism*; D007562:Creutzfeldt-Jakob Syndrome / Q000378:metabolism*; D003666:Decontamination / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D018620:PrPSc Proteins / Q000187:drug effects / Q000378:metabolism*; D011994:Recombinant Proteins / Q000187:drug effects / Q000378:metabolism; D012972:Sodium Hydroxide / Q000494:pharmacology; D013193:Stainless Steel; D013232:Steel; D013499:Surface Properties; D013525:Surgical Instruments
D000682:Amyloid; D018620:PrPSc Proteins; D011994:Recombinant Proteins; D013193:Stainless Steel; D013232:Steel; D012972:Sodium Hydroxide