An advantageous method utilizing new homogenizing device BioMasher and a sensitive ELISA to detect bovine spongiform encephalopathy accurately in brain tissue.
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.
Yamamoto
Takuji
T
Nippi Research Institute of Biomatrix, 520-11 Kuwabara, Toride-shi, Ibaraki 302-0017, Japan.
Ushiki
Yuko
Y
Hara
Susumu
S
Hall
William W
WW
Tsukagoshi-Nagai
Hiroe
H
Yokoyama
Takashi
T
Tagawa
Yuichi
Y
Sata
Tetsutaro
T
Yamakawa
Yoshio
Y
Kinoshita
Noriaki
N
Tamura
Fjio
F
Hattori
Shunji
S
Irie
Shinkichi
S
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
D000911:Antibodies, Monoclonal; D011328:Prions