title: Failure of immunocompetitive capillary electrophoresis assay to detect disease-specific prion protein in buffy coat from humans and chimpanzees with Creutzfeldt-Jakob disease.
abstract: The emergence of a new environmentally caused variant of Creutzfeldt-Jakob disease (vCJD), the result of food-born infection by the causative agent of bovine spongiform encephalopathy (BSE), has stimulated research on a practical diagnostic screening test. The immunocompetitive capillary electrophoresis (ICCE) assay has been reported to detect disease-specific, proteinase-resistant prion protein (PrPres) in the blood of scrapie-infected sheep. We have applied this method to blood from CJD-infected chimpanzees and humans. The threshold of detection achieved with our ICCE was 0.6 nM of synthetic peptide corresponding to the prion protein (PrP) C-terminus, and 2 nM of recombinant human PrP at the optimized conditions. However, the test was unable to distinguish between extracts of leucocytes from healthy and CJD-infected chimpanzees, and from healthy human donors and patients affected with various forms of CJD. Thus, the ICCE assay as presently performed is not suitable for use as a screening test in human transmissible spongiform encephalopathies (TSEs).
authors.affiliation: Plasma Derivatives Department, Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross, Rockville, MD 20855, USA. cervenak@usa.redcross.org
authors.forename: Larisa
authors.initials: L
authors.lastname: Cervenakova
authors.forename: Paul
authors.initials: P
authors.lastname: Brown
authors.forename: Serguei
authors.initials: S
authors.lastname: Soukharev
authors.forename: Oksana
authors.initials: O
authors.lastname: Yakovleva
authors.forename: Heino
authors.initials: H
authors.lastname: Diringer
authors.forename: Evgueni L
authors.initials: EL
authors.lastname: Saenko
authors.forename: William N
authors.initials: WN
authors.lastname: Drohan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001666:Binding Sites, Antibody; D007562:Creutzfeldt-Jakob Syndrome / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D019075:Electrophoresis, Capillary / Q000379:methods*; D005069:Evaluation Studies as Topic; D006801:Humans; D007962:Leukocytes / Q000737:chemistry; D008969:Molecular Sequence Data; D009154:Mutation; D002679:Pan troglodytes; D010455:Peptides / Q000032:analysis / Q000276:immunology; D018620:PrPSc Proteins / Q000032:analysis / Q000097:blood* / Q000235:genetics
chemical_list: D000906:Antibodies; D010455:Peptides; D018620:PrPSc Proteins