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Effect of Tissue Deterioration on Postmortem BSE Diagnosis by
Immunobiochemical Detection of an Abnormal Isoform of Prion Protein
Hiroko HAYASHI1), Masuhiro TAKATA1), Yoshifumi IWAMARU1), Yuko
USHIKI1)2), Kumiko M. KIMURA1), Yuichi TAGAWA1), Morikazu SHINAGAWA1)
and Takashi YOKOYAMA1)
1) Prion Disease Research Center, National Institute of Animal Health
2) Nippi Research Institute of Biomatrix
(Received 25-Aug-2003)
(Accepted 14-Jan-2004)
ABSTRACT. 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 (PrPSc) 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 PrPSc 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 PrPSc was observed in WB, even when
performed after 4 days of incubation at 37°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 PrPSc 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.
KEY WORDS: BSE, ELISA, PrPSc, Western blot
[PDF (96K) ]
[References ]
To cite this article:
Hiroko HAYASHI, Masuhiro TAKATA, Yoshifumi IWAMARU, Yuko USHIKI, Kumiko
M. KIMURA, Yuichi TAGAWA, Morikazu SHINAGAWA and Takashi YOKOYAMA
Effect of Tissue Deterioration on Postmortem BSE Diagnosis by
Immunobiochemical Detection of an Abnormal Isoform of Prion Protein. J.
Vet. Med. Sci.. Vol. 66: 515-520. (2004) .
------------------------------------------------------------------------
DOI 10.1292/jvms.66.515
JOI JST.JSTAGE/jvms/66.515
Copyright(c) 2004 by the Japanese Society of Veterinary Science
Immunobiochemical Detection of an Abnormal Isoform of Prion Protein
Hiroko HAYASHI1), Masuhiro TAKATA1), Yoshifumi IWAMARU1), Yuko
USHIKI1)2), Kumiko M. KIMURA1), Yuichi TAGAWA1), Morikazu SHINAGAWA1)
and Takashi YOKOYAMA1)
1) Prion Disease Research Center, National Institute of Animal Health
2) Nippi Research Institute of Biomatrix
(Received 25-Aug-2003)
(Accepted 14-Jan-2004)
ABSTRACT. 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 (PrPSc) 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 PrPSc 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 PrPSc was observed in WB, even when
performed after 4 days of incubation at 37°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 PrPSc 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.
KEY WORDS: BSE, ELISA, PrPSc, Western blot
[PDF (96K) ]
[References ]
To cite this article:
Hiroko HAYASHI, Masuhiro TAKATA, Yoshifumi IWAMARU, Yuko USHIKI, Kumiko
M. KIMURA, Yuichi TAGAWA, Morikazu SHINAGAWA and Takashi YOKOYAMA
Effect of Tissue Deterioration on Postmortem BSE Diagnosis by
Immunobiochemical Detection of an Abnormal Isoform of Prion Protein. J.
Vet. Med. Sci.. Vol. 66: 515-520. (2004) .
------------------------------------------------------------------------
DOI 10.1292/jvms.66.515
JOI JST.JSTAGE/jvms/66.515
Copyright(c) 2004 by the Japanese Society of Veterinary Science
