Specific detection of prion antigenic determinants retained in bovine meat and bone meal by flow microbead immunoassay.
OBJECTIVE
The purpose of this study was to develop an effective method for detecting prion (PrP) antigenic determinants remaining in bovine meat and bone meal (MBM) using pressurized fluid extraction (PSE) equipment and flow microbead immunoassay (FMI).


RESULTS
Using the FMI, bovine recombinant PrP could be determined quantitatively in the 7 pmol-7 nmol range using anti-PrP peptide polyclonal antibody-coupled microbeads and anti-PrP monoclonal antibody (SAF61) as a detection antibody. PSE extraction at 120 degrees C for 5 min under high pressure was most effective for eluting PrP determinants from bovine MBMs. The FMI was capable of detecting PrP determinants in bovine MBM extracts with high specificity and indicated that the MBMs contained high levels of PrP determinants. This assay was also applied to the detection of PrP(Sc) determinants in bovine MBM spiked with a scrapie-infected brain at a weight ratio of 50 : 1.


CONCLUSIONS
These data indicate that this assay was effective for the specific detection of PrP determinants contained in bovine MBM extracts.


CONCLUSIONS
To our knowledge, this is the first report detailing the detection of PrP determinants in bovine MBM. The assay could be applied to securing the safety of bovine MBM.
Murayama
Y
Y
Prion Disease Research Center, National Institute of Animal Health, Ibaraki, Japan. ymura@affrc.go.jp
Yoshioka
M
M
Horii
H
H
Takata
M
M
Miura
K
K
Shinagawa
M
M
D000821:Animal Feed; D000818:Animals; D000918:Antibody Specificity; D001688:Biological Products / Q000737:chemistry; D015153:Blotting, Western / Q000379:methods; D002417:Cattle; D000939:Epitopes / Q000032:analysis*; D017022:Flow Injection Analysis; D005506:Food Contamination; D007118:Immunoassay / Q000379:methods; D008460:Meat / Q000032:analysis*; D008863:Microspheres; D008903:Minerals / Q000737:chemistry; D018620:PrPSc Proteins / Q000276:immunology*; D012608:Scrapie / Q000276:immunology; D012756:Sheep
D001688:Biological Products; D000939:Epitopes; D008903:Minerals; D018620:PrPSc Proteins; C027762:bone meal