title: An advantageous method utilizing new homogenizing device BioMasher and a sensitive ELISA to detect bovine spongiform encephalopathy accurately in brain tissue.
abstract: A new screening method was developed to detect bovine spongiform encephalopathy (BSE). This method is advantageous because it has a simpler and safer protocol than commercial kits. A new device was developed for this method; it was named the BioMasher, to homogenize brain tissue by passing it through a porous rigid polypropylene filter. In this system, a purification step was eliminated in the sample preparation. Thus, the time needed for sample pretreatment is substantially shortened, and the risk of infection during sample processing is effectively reduced. Monoclonal antibodies to prion protein were created and used to construct a sensitive sandwich enzyme-linked immunosorbent assay system. The sensitivity of this assay kit using frozen BSE-positive brain is comparable or more sensitive than commercial kits. Moreover, the detection sensitivity for deteriorated samples, which were kept at 37 degrees C for 1 day, is 10- to 30-fold more sensitive than a commercial kit.
authors.affiliation: Nippi Research Institute of Biomatrix, 520-11 Kuwabara, Toride-shi, Ibaraki 302-0017, Japan.
authors.forename: Takuji
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Yuko
authors.initials: Y
authors.lastname: Ushiki
authors.forename: Susumu
authors.initials: S
authors.lastname: Hara
authors.forename: William W
authors.initials: WW
authors.lastname: Hall
authors.forename: Hiroe
authors.initials: H
authors.lastname: Tsukagoshi-Nagai
authors.forename: Takashi
authors.initials: T
authors.lastname: Yokoyama
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Tagawa
authors.forename: Tetsutaro
authors.initials: T
authors.lastname: Sata
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Yamakawa
authors.forename: Noriaki
authors.initials: N
authors.lastname: Kinoshita
authors.forename: Fjio
authors.initials: F
authors.lastname: Tamura
authors.forename: Shunji
authors.initials: S
authors.lastname: Hattori
authors.forename: Shinkichi
authors.initials: S
authors.lastname: Irie
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001923:Brain Chemistry; D002417:Cattle; D016643:Encephalopathy, Bovine Spongiform / Q000175:diagnosis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005374:Filtration / Q000379:methods; D051379:Mice; D011328:Prions / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D011328:Prions