Effect of tissue deterioration on postmortem BSE diagnosis by immunobiochemical detection of an abnormal isoform of prion protein.
Surveillance for bovine spongiform encephalopathy (BSE) in fallen stock in Japan is conducted with a commercial enzyme-linked immunosorbent assay (ELISA) for mass screening, with Western blotting (WB) and immunohistochemistry performed for confirmation of the ELISA. All tests are based on immunological detection of an abnormal isoform of the prion protein (PrP(Sc)) in brain tissues, which have sometimes deteriorated by the time samples from fallen stock reach a diagnostic laboratory. To evaluate BSE surveillance procedures for fallen stock, we examined PrP(Sc) detection from artificially deteriorated BSE-affected bovine brain tissues with a commercial ELISA kit and compared the results with those of WB. The optical density (OD) values of the ELISA decreased with advancing deterioration of the tissues, whereas no reduction in the signal for PrP(Sc) was observed in WB, even when performed after 4 days of incubation at 37 degrees C. The progressive decrease in the OD values in the ELISA appear to be caused by a partial loss of the N-terminal moiety of PrP(Sc) due to digestion by endogeneous and/or contaminated microbial enzymes, and by the presence of ELISA inhibitors that are generated in deteriorated tissues. These results suggest that WB is the most reliable test for fallen stock, especially for cattle brains within decaying carcasses.
Hayashi
Hiroko
H
Prion Disease Research Center, National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
Takata
Masuhiro
M
Iwamaru
Yoshifumi
Y
Ushiki
Yuko
Y
Kimura
Kumiko M
KM
Tagawa
Yuichi
Y
Shinagawa
Morikazu
M
Yokoyama
Takashi
T
D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D001921:Brain / Q000473:pathology*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000473:pathology; D016643:Encephalopathy, Bovine Spongiform / Q000175:diagnosis* / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D007150:Immunohistochemistry / Q000662:veterinary; D011180:Postmortem Changes; D011328:Prions / Q000302:isolation & purification*
D011328:Prions